3

Prospectus 2018 Institute for Protein Research Osaka University

OSAKA UNIVERSITY

IPR
Prospectus 2018

Institute
I tit t for
f Protein
P t i ResearchRhOsaka University

IPR

Institute for Protein Research Osaka University

CONTENTS
Message from the Director

1

Concept and Future2

Organization3

Sixty-year history of IPR

4

Activities 2017-2018

6

Division of Protein Chemistry

9

Laboratory of Protein Organic Chemistry
Laboratory of Cell Systems
Laboratory of Nanobiology
Laboratory of Organelle Biology

Division of Protein Structural Biology

17

Laboratory of Protein Folding
Laboratory of Molecular Biophysics
Laboratory of Protein Crystallography
Laboratory of Membrane Protein Chemistry

Division of Integrated Protein Functions

25

Laboratory of Genome and Chromosome Functions
Laboratory for Molecular and Developmental Biology
Laboratory for Advanced Brain Functions
Laboratory of Homeostatic Integration

Division of Multiscale Integrative Protein Science

33

Laboratory of Nuclear Network
Laboratory of Multiscale Structural Biology
Laboratory for Infection Systems
Laboratory of Visiting Scientists

Research Center for State-of-the-Art Functional Protein Analysis

39

Laboratory of Protein Profiling and Functional Proteomics
Laboratory of Protein Synthesis and Expression
Laboratory of Supramolecular Crystallography
Laboratory of Advanced Protein Characterization
Laboratory of Protein Databases
Open Space Laboratory for Advanced Protein Science

Technology Division

53

Technology Division

Division of Donated Fund Research

55

Division of Matrixome Research and Application

About IPR

57

Educational Activities
Research Activities

Public Relations Office

Joint-Usage / Research Center
IPR Saff

Institute for Protein Research, Osaka University

Administrative Council

3−2 Yamadaoka, Suita Osaka, 5650871, Japan

Staff and Accounting Data

http://www.protein.osaka-u.ac.jp

Floor PlanACCESS MAP

68

FROM THE DIRECTOR

Message from the Director

Message

Institute for Protein Research Osaka University

In 2018, the Institute for Protein Research (IPR), Osaka
University celebrates the 60 th anniversary since its foundation. On behalf of all the members of IPR, I would like toexpress my sincere appreciation for researchers both fromJapan and overseas who have collaborated with researchers at IPR, government oﬃcials who have supported ouractivities, as well as academic, technical and administrative
staffs, and students who have contributed to the development of IPR.
From the very beginning of its foundation in 1958, researchers in the fields of chemistry, biology, physics, andmedicine gathered at IPR to pursue interdisciplinary studies
of proteins. Since that time, the IPR has been at the cuttingedge of new developments in protein science aimed atachieving an understanding of the mechanism of living systems – from the molecular level to organism level.
The IPR started with only three divisions (laboratories): Organic Chemistry, Physical Chemistry and Protein Metabolism. After sixty years of steady development, the IPR now
consists of five divisions (17 laboratories) with an attachedcenter, the Research Center for State-of-the-Art Functional
Protein Analysis (6 laboratories), which aims to develop andapply original techniques to the structural and functional
studies of proteins.
In April 2010, the IPR was designated as one of the Joint
Usage/Research Centers in Japan by the Ministry of Education, Culture, Sports, Science and Technology (MEXT).
In regard to its role as a Joint Usage/Research Center, theIPR offers three collaborative opportunities to help advancejoint research activities in the protein science field. The IPR
also manages access to a range of high performanceequipment, such as the“Synchrotron Beamline at SPring-8”,
the“High Magnetic Field Nuclear Magnetic Resonance(NMR) spectrometers”and the“Advanced Cryo-Electron Microscopy Suite”, all of which are open to protein science
researchers both in Japan and overseas. In addition, as oneof the four members of the wwPDB (worldwide Protein Data
Bank), the IPR operates the PDBj (Protein Data Bank Japan).
The PDBj-BMRB also constructs and manages an NMR
experimental database, in collaboration with the BMRB
(BioMagResBank) in the USA. The IPR has also extensively organized a very large number of diverse international

collaborative research projects with overseas protein scientists and institutions under the auspices of the “International Collaboration Research Program”.
Total of more than 60 academic staff at the IPR work very
hard for their own scientific research, as well as for theireducational activities directed towards teaching Osaka University students at the undergraduate level (placed within
the Faculties of Science, and Medicine) and Master’scourse and Doctor’s course students (placed within the
Graduate Schools of Science, Medicine, and Frontier Biosciences). From those faculties and graduate schools, about
130 students study in the laboratories at IPR. In addition tothis student cohort, about 40 postdoctoral fellows maketheir own original investigations in support of various national and international funded research project. We regard
these young scientists, who come to the IPR from countries
all over the world, as important human resources of the IPR
and they are expected to lead new protein science fields inthe future.
Recently, the life science field has made dramatic developments. We are now on the cusp of developing a new proteinscience field that is capable of revealing the secrets of higher-order living systems through investigations of the interactions occurring between their networks of biological molecules. These functional studies are facilitated by knowledgeof the precise atomic structures of biological molecules. To
actualize this future potential, we aim to develop and combine a very large array of different techniques. With this IPR
research cohort, we are in position to strive towards thedevelopment of this new type of protein science – one that
represents a future approach to the life sciences based on
the eradication of borders between laboratories, scientificfields, facilities and indeed countries, and most importantly,
one that is capable of developing both the scientific breakthroughs and the important human resources of the next
generation.
I hope that the IPR Prospectus 2018 well demonstrates ourresearch and activities, as well as our continued efforts fordevelopment.
Atsushi Nakagawa, D.Sci., Director◦◦◦ 1 ◦

IPR
Concept and Future
The processes of life, mediated by a large variety of3) IPR will educate undergraduate/graduate students, Ph.

proteins, are very complex. Elucidation of these processes

D. students, and young researchers not only in Osaka

through molecular studies of proteins can be achieved by

University but also other universities including overseas

efficient cooperation of researchers in various fields ofusing the framework of the joint-use/research center.

natural sciences. Another requisite is the establishment of

Based on the above mission, IPR has so far initiatedsuitable infrastructure that allows close collaborations

various projects on structural biology, neurobiology,

among scientists across the country. The Institute forproteomics, and molecular and cellular biology, andProtein Research (IPR) was founded as a joint-use

produced many excellent results. Although IPR started as

research organization attached to Osaka University toa domestic center, it is now widely recognized as anfulfill these needs, and thus to play a central role in protein

international center of excellence for protein research. For

science in Japan. Now, IPR works as one of the Joint

instance, IPR operates the Worldwide Protein Data Bank

Usage/Research(wwPDB) as one of four worldwide centers, mainlyCenters

authorized

byJapanese

Government.

covering the Asia region. Because of its high standard ofThus, the mission of IPR has not changed during its longscientific activity, it has attracted many researchers from

history, and it is summarized as the following threeabroad and will continue to do so in the future. In April

concepts:

2016, a new program started to aim for the international

1) Basic studies are performed on the structure andcore for the multiscale integrative structural life science.

function of proteins and their biological significance at

IPR also invites researchers in the new research fields

the molecular level, as well as at the cellular level withsuch as systems biology and single-molecule analysis. In

tight collaborations by researchers in IPR having

addition, two new divisions of donated fund research also

various backgrounds such as chemistry, physics,

started their activities with the associated industries inbiology and medicine.

2016.

2) IPR provides the resources for research, databases
such as PDB, and scientific communication for

The

IPR

will

continuetomakecontributions to revealing the structure and function ofproteins and their networks for the elucidation of life.

domestic and international collaborative studies as the
Joint Usage/Research Center, thereby promoting
protein science for the scientific community andsociety.

Foreign researchers
and institutions

Division ofProtein Chemistry
4 PI laboratories

Division of Protein
Structural Biology
4 PI laboratories
Division of Multiscale
Integrative Protein Science
4 PI laboratories

Research Center for State-of-the-Art
Functional Protein Analysis
6 PI laboratories
Division of Donated Fund Research
Nippi Matrixone
Research and Applications

Industries andCommunity◦ 2 ◦◦◦

essential

Domestic researchers
and institutions

Division ofIntegrated Protein Functions
4 PI laboratories

Institute for Protein Research Osaka University

Organization
Director
Faculty Meeting
Administrative Council
Panel on Joint Usage/Research
Research Divisions
Division of Protein Chemistry
Laboratory of Protein Organic Chemistry
Laboratory of Cell Systems
Laboratory of Nanobiology
Laboratory of Organelle Biology

Division of Protein Structural Biology
Laboratory of Protein Folding
Laboratory of Molecular Biophysics
Laboratory of Protein Crystallography
Laboratory of Membrane Protein Chemistry

Division of Integrated Protein Functions
Laboratory of Genome and Chromosome Functions
Laboratory for Molecular and Developmental Biology
Laboratory for Advanced Brain Functions
Laboratory for Nuclear Dynamics
Laboratory of Homeostatic Integration

Division of Multiscale Integrative Protein Science
Laboratory of Nuclear Network
Laboratory of Multiscale Structural Biology
Laboratory for Infection Systems
Laboratory of Visiting Scientists

Special Research Facilities
Research Center for State-of-the-Art Functional Protein Analysis
Laboratory of Protein Profiling and Functional Proteomics
Laboratory of Protein Synthesis and Expression
Laboratory of Supramolecular Crystallography
Laboratory of Advanced Protein Characterization
Laboratory of Protein Databases
Open Space Laboratory for Advanced Protein Science

Division of Donated Fund Research
Division of Matrixome Research and Application

Technology Division
Administrative Office
General Affairs Section
Library
Accounting Section
Project Team of Joint Usage / Research Center
Research Support Section

Public Relations Office
IPR Alumni Office◦◦◦ 3 ◦

IPR
Sixty-year history of IPR
Osaka University has been active in the study of proteinsand enzymes since its foundation in 1931, and it was along-standing desire of the university authorities to further
promote this facet of the university’s activities by
establishing a research institute specialized for proteinscience. In 1955, an oﬃcial plan was drafted to establish
such an institute as a part of Osaka University andsubmitted to the Ministry of Education, Science andCulture. The Ministry agreed to open a new laboratory fororganic chemistry of proteins and amino acids in the
Faculty of Science. The new laboratory was opened in
1956, and Professor Shiro Akabori, who had played apivotal role in protein research, was appointed as its
supervisor.
In the meantime, a strong desire for the establishment ofa central institute for protein research with the aim offacilitating close cooperation among researchers from awide variety of scientific fields had continuously grownamong scientists in the relevant fields. In 1957, the
Science Council of Japan urged the Government toconsider the foundation of such an institute somewhere in
the country. The Government decided to establish a
research institute for protein science attached to Osaka
University. The Institute for Protein Research (IPR) was
thus formally founded on April 1, 1958, as a part of Osaka
University, and Professor Shiro Akabori was appointed asits first director. IPR has developed significantly in termsof both scientific activity and infrastructure. In 2016, a
new division of Multiscale Integrative Protein Science andtwo divisions of donated fund research have been founded.
Now, the IPR comprises four divisions with 17 laboratories,
an attached research center (Research Center for Stateof-the-Art Functional Protein Analysis) with 6 laboratories,
and one division of donated fund research, serving as a
joint usage/research institute for the community in thefields of protein and related sciences.

The Research Center for Protein Engineering was established.
Suita Campus（1972）1988
Buildings for the Crystallographic Research Center(1,505 ㎡) and NMR Research Laboratory (267㎡)
were completed.

◦ The 5th Director

Masao Kakudo

1961◦ The 1st Director

Shiro Akabori

Faculty of Science◦ 4 ◦◦◦

1972

The crystal structure of bonito heart ferrocytochrome c was determinedby Prof. Kakudo of IPR for the first time in Japan.
Main building (7,873 ㎡)of the IPR was completed
on Suita Campus.

1956

1958

1979

1971
Main building (4,130㎡）was completed on the former
Nakanoshima Campus.
Establishment of the Institute for Protein Research
as a Joint-use Research Organization, composedof three Divisions : Organic Chemistry, Physical
Chemistry and Protein Metabolism

Set up of a new laboratory in Faculty of Science, Osaka University, fororganic chemical studies of proteins and amino acids (the predecessor of
IPR) supervised by Prof. Shiro Akabori.

The first IPRbuilding

Institute for Protein Research Osaka University

The 60th Anniversary

2018

Certified continuous activities asa Joint Usage/Research Center by MEXT
The Research Center for State-of-the-Art
Functional Protein Analysis was established.

2009
2008

2004
2007 Retreat

2002
2001
1998

2012

2010

Certified as a Joint Usage/Research Center by MEXT.
Main building reinforcement forearthquake-resistance was completed.

2016

The 50th Anniversary
Building of Collaborative Research
Facility (1,149㎡) was completed.

Transformed into a Research Institute ofJapanese National Universities under the
National University Corporation Law

The Research Center for Structural andFunctional Proteomics was established.

The Protein Data Bank Japan（PDBj）
started its operation.
The Center for Structural
Biology was established.
Suita Campus（2008）

Development of the Attached Research Centers
Peptide Center（1962）
Crystallographic Research Center（1978）

Professors Emeriti
Akira Asano, D.Sci.
Toshio Takagi, D.Sci.

Research Center for Protein Engineering（1988）
Center for Structural Biology（1998）
Research Center for Structural and Functional Proteomics（2002）
Reseach Center for State-of-the-Art Functional Protein Analysis（2012）

Fumio Sakiyama, D.Sci.
Katsuya Nagai, M.D., D.Med.
Hideo Akutsu, D.Sci.
Tomitake Tsukihara, D.Sci.
Saburo Aimoto, D.Sci.
Kiyotoshi Sekiguchi, D.Sci.
Shoji Tajima, D.Sci.

Nakanoshima Campus（1961）

Toshiharu Hase, D.Sci.
Kazuaki Yoshikawa,M.D., D.Med.

Opening of the IPR（1961）

Haruki Nakamura, D.Sci.

Directors
1st Shiro Akabori

Apr. 1, 1958 − Nov.30,1961

2nd Toshizo Isemura

Dec. 1, 1961 − Nov.30,1965

3rd Tomoji Suzuki

Dec. 1, 1965 − Aug.14,1969

4th Kouzo Narita

Aug.15, 1969 - Aug.14,1971

5th Masao Kakudo

Aug.15, 1971 − Apr. 1,1982

6th Yoshiharu Izumi

Apr. 2, 1982 − Mar.31,1985

7th Ryo Sato

Apr. 1, 1985 − Mar.31,1987

8th Takekazu Horio

Apr. 1, 1987 − Mar.31,1989

9th Yukiteru Katsube

Apr. 1, 1989 − Mar.31,1993

10th Hachiro Nakagawa

Apr. 1, 1993 − Mar.31,1995

11th Fumio Sakiyama

Apr. 1, 1995 − Mar.31,1997

12th Yoshimasa Kyogoku

Apr. 1, 1997 − Mar.31,1999

13th Yasutsugu Shimonishi Apr. 1, 1999 − Mar.31,2000
14th Katsuya Nagai

Apr. 1, 2000 − Mar.31,2004

15th Hideo Akutsu

Apr. 1, 2004 − Mar.31,2006

16th Tomitake Tsukihara

Apr. 1, 2006 − Mar.31,2008

17th Saburo Aimoto

Apr. 1, 2008 − Mar.31,2010

18th Toshiharu Hase

Apr. 1, 2010 − Mar.31,2014

19th Haruki Nakamura

Apr. 1, 2014 − Mar.31,2018

20th Atsushi Nakagawa

Apr. 1, 2018 − Present◦◦◦ 5 ◦

IPR
Activities 2017 − 2018
IPR Seminar

IPR holds IPR Seminars on various topics throughout the year as one of the seven collaborative activities of Join-Usage/
Research Center.

Title

August 24, 2017 at Lecture Hall of IPR
Future of Hyper-Polarized Nuclear Spins-State-of-the-Art DNP Techniques-

January 5, 2018 at Seoul National University“Japan-Korea Bilateral Symposium between SNU
and IPR on Structure and Folding of Disease Related
Proteins”

February 20, 2018 at Icho Kaikan, Suita Campus
PDBj & BINDS Joint Workshop “Introduction to 3D
macromolecular data for life science”◦ 6 ◦◦◦

Date

Venue

2017/5/12-14

SPring-8

2017/6/21

Sendai International
Center1

Workshop on the Beamline 20172

Toward International Cooperation in Protein Science3

The 2nd Molecular and Cellular Life Sciences (MCLS2017):
Structural biology, modeling and molecular dynamics withapplications in biotechnology and medicine

2017/7/17-18

Wyndham Hotel,
Surabaya, Indonesia

4

Frontiers of interdisciplinary research on membrane iontransport: from computational science to medicine

2017/7/27-28

IPR

5Current status and future plan in protein structure biology
research at SPring-8

2017/8/3-4

IPR

6

Progress and Future of Protein Crystallography in Japan In Memory of Professor Katsube

2017/8/4

IPR

7

Future of Hyper-Polarized Nuclear Spinsー State-of-the-Art DNP Techniques ー2017/8/24-25

IPR

8

The 3rd Trilateral Workshop for Frontier Protein Studies

2017/8/31-9/2

National Center forProtein Science,
Shanghai, China

9

Protein solution research geared toward industrial application

2017/9/11-12

IPR

10

Synthesis and functional analysis of biomolecule containing
chalcogen and heteroatom

2017/11/1

IPR

11Australian National University (ANU) &IPR 2nd Joint
Symposium 2017 "Protein Structure and Function"

2017/12/3-5

IPR

12

Establishment of Structural Biology Network in Asia andOceania

2017/12/6-7

National Synchrotron
Radiation ResearchCenter, Taiwan

13Japan-Korea Bilateral Symposium between SNU and IPR on
Structure and Folding of Disease Related Proteins2018/1/5

College of Pharmacy,
Seoul National
University, Korea

14

Frontiers in vision and sensation research

2018/1/19-20

Senri Life Science
Center

15

PDBj & BINDS Joint Workshop“Introduction to 3D
macromolecular data for life science”

2018/2/20

Icho Kaikan,
Osaka Univ.

16

Now and future in next generation antibodies
ー from basic research to drug development －2018/2/22

Asano Campus,
Tokyo Univ.

17

Mechanism of Biology on the Membrane

2018/3/1-2

IPR

18

Symposium on Chromosome Biology and Cell Signalling(Jointly organized by IISER TVM and Osaka University)

2018/3/5

Indian Institute of
Science, India

19

Future of mass spectrometry

2018/3/9-10

Graduate School of
Science, Osaka Univ.

20

New Frontiers in Molecular Membrane Biology using in vitro
Reconstitution Approaches

2018/3/27

IPR

Institute for Protein Research Osaka University

IPR Retreat 2017 December 12-13, 2017

Icho Kaikan, Suita Campus, Osaka University

IPR holds a retreat every year with the aim to facilitate

as a seminar session by three lecturers invited from othercommunication and interaction among researchers andresearch institutes. It was a great opportunity for students

students at IPR. The Retreat 2017 consisted of six

and academic staff at IPR to discuss and deepen

workshops in the fields of cryo-EM, bioinformatics, peptide

understanding of research activities conducted at IPR.

synthesis, systems biology, and molecular imaging, as well

IPR Colloquia

As a measure for faculty development, IPR holds colloquia

five times each year. Young researchers are selected to
be speakers, which provides them with opportunities forpresentation and discussion with other researchers. The
IPR Colloquia has been a forum for active interaction andfuture collaboration in IPR.

Outreach Activities

IPR has actively promoted outreach activities with the aimof promoting science to the public and thus contribute tosociety. Academic staff of IPR has provided lectures forjunior high school/high school students as well as thegeneral public; and has participated in science events
such as JST Science Agora held annually in Tokyo.
JST Science Agora 2017
November. 24-26, 2017
Telecom Center Building, Tokyo

Osaka Univ. Icho Festival 2018
April. 30, 2018
Institute for Protein Research◦◦◦ 7 ◦

IPR
International Exchange

IPR has been actively promoting international exchangeorative Research program, for overseas researchers who

through various programs.

perform coordinated researches at the institute and use

The Research Center for State-of-the-Art Functional Pro-

particular experimental facilities of IPR including the syn-tein Analysis of IPR has provided positions for visiting pro-chrotron beamline for biological macromolecular assem-

fessors from overseas. IPR accepts international scientists

blies at SPring-8. IPR has also been accepting internation-

through programs sponsored by the Japan Society for theal exchange students through a program called FrontierLab@

Promotion of Science (JSPS) and other programs when fi-

OsakaU.

nancing is guaranteed from outside IPR. More than two

To promote further cooperation in research, IPR has con-hundred visiting international scientists have participated included Inter-Faculty Academic Exchange Agreements withresearch activities since the establishment of the IPR. In

overseas institutes as listed below:

2005, IPR started a new program, the International Collab-

Inter-Faculty Academic Exchange Agreements
Institution

Country

Year ofagreement

1

Center for Genetic Engineering and Biotechnology

Cuba

2003

2

National Synchrotron Radiation Research Center

Taiwan2007

3

Indian Institute of Chemical Biology

India

2009

4

Institute of Tropical Disease, Airlangga University

Indonesia

20145

National Center for Protein Science Shanghai

China

2014

6

Center of Protein Science, Peking University

China

2014

Australia

2014

Korea

2015

USA

2015

Taiwan

2015

7
8
9
10

College of Physical and Mathematical Sciences,
Australian National University
College of Pharmacy, Seoul National University
Center for Integrative Proteomics Research, The
State University of New Jersey, Rutgers
College of Life Science, National Tsing Hua Univ
ersity11

Panjab University

India

2017

12

University College Dublin

Ireland

2017

India

2017

USA

2017

Germany

2017

13
14
15

Indian Institute of Science Education and Research (IISERs) Thiruvananthapuram
University of Chicago
Faculty of Biology and Biotechnology, Ruhr University Bochum (RUB)

May 9 & 10, 2018 Bilateral Symposium &Workshop between Astbury Center of Leeds and IPR◦ 8 ◦◦◦

IPR’s Global Projects page on IPR website
http://www.protein.osaka-u.ac.jp/Global/in
dex.html

October 2017
A cooperation agreement
has been concluded
between the Faculty ofBiology and Biotechnology,
Ruhr University Bochum(RUB) and IPR.

Division of Protein
Chemistry

Laboratory of Protein Organic Chemistry
Laboratory of Cell Systems
Laboratory of Nanobiology
Laboratory of Organelle Biology

IPR
㻌

/DERUDWRU\ RI 3URWHLQ 2UJDQLF &KHPLVWU\
㻌/DERUDWRU\RI3URWHLQ2UJDQLF&KHPLVWU\
㻌㻌㻌
3URIHVVRU+LURQREX+RMR3K'
KRMR#SURWHLQRVDNDXDFMS $VVRFLDWH3URIHVVRU

7RUX.DZDNDPL3K'$VVLVWDQW3URIHVVRU<X\D$VDKLQD3K'

 &KHPLFDOV\QWKHVLVHQDEOHVWKHSUHSDUDWLRQRISURWHLQVZKLFKDUHGLIILFXOWWRSUHSDUHE\WKHUHFRPELQDQWPHWKRG VXFK DV VLWHVSHFLILFDOO\ ODEHOHG JO\FRV\ODWHG DQGSKRVSKRU\ODWHG SURWHLQV 7KH /DERUDWRU\ RI
3URWHLQ2UJDQLF&KHPLVWU\LVDLPLQJWRSURPRWHQHZSURWHLQUHVHDUFKHVXVLQJWKHVHV\QWKHWLFSURWHLQV

7KXVRXUODERUDWRU\LVGHYHORSLQJIDFLOHPHWKRGVIRUSURWHLQV\QWKHVLVEDVHGRQOLJDWLRQFKHPLVWULHV$W

SUHVHQWZHDUHDLPLQJWRRYHUFRPHDVROXELOLW\SUREOHPZKLFKZHRIWHQIDFHGXULQJWKHOLJDWLRQUHDFWLRQ
7KHSUREOHPLVSDUWLFXODUO\VHULRXVIRUWKHV\QWKHVLVRIPHPEUDQHSURWHLQV7RRYHUFRPHWKLVSUREOHPZH

DUH V\QWKHVL]LQJ PHPEUDQH SURWHLQV DQG WKHLU SDUWLDO VHTXHQFHV WR HOXFLGDWH WKH VLJQDO WUDQVGXFWLRQPHFKDQLVP E\ VROLG VWDWH 105 DQG ,5 0RGLILHG KLVWRQHV DQG WKHLU SDUWLDO VHTXHQFHV JO\FRV\ODWHG

SURWHLQVDUHDOVREHLQJV\QWKHVL]HGIRUIXQFWLRQDODQDO\VHV

5HVHDUFK3URJUDPV*HQHUDOVWXGLHVRQDFKHPLFDOSURWHLQV\QWKHVLV

6\QWKHWLFVWXGLHVRIPRGLILHGKLVWRQHVDQGHOXFLGDWLRQRIWKHUROHRIPRGLILHV

6\QWKHVLVRIPHPEUDQHSURWHLQVDQGWKHLUVWUXFWXUDODQGIXQFWLRQDOVWXGLHV6\QWKHVLVRIJO\FRSURWHLQVDQGWKHLUIXQFWLRQDOVWXGLHV

5HVHDUFK7RSLFV‘ /LJDWLRQFKHPLVWULHVKDYHEHHQZLGHO\XVHGIRUWKHV\QWKHVLVRI JO\FR  SURWHLQV IRU WKHLU IXQFWLRQDO DQG VWUXFWXUDO VWXGLHV+RZHYHU ZH RIWHQ IDFH D VHULRXV VROXELOLW\ SUREOHP GXULQJV\QWKHVLV E\ OLJDWLRQ PHWKRGV HYHQ LQ WKH FDVH RI JOREXODU
SURWHLQV7KHVDPHSUREOHPDOVRRFFXUVGXULQJWKHV\QWKHVLVRIKLJKO\K\GURSKRELFSURWHLQVVXFKDVPHPEUDQHSURWHLQVZKLFKDUH LPSRUWDQW GUXJ WDUJHWV 7R RYHUFRPH WKLV SUREOHP ZH
H[DPLQHG WKH PRGLILFDWLRQ RI WKH VWUXFWXUH RI SRO\SHSWLGHV
GXULQJ WKH V\QWKHVLV ,Q WKH FDVH RI KXPDQ LQWHUOHXNLQ IRU
H[DPSOHWKHLQWURGXFWLRQRIEDVLFSLFRO\OHVWHUZDVLQWURGXFHG
WRWKHVLGHFKDLQFDUER[\JURXSRI*OXUHVLGXHZKLFKUHYHUVHVWKH SRODULW\ IURP QHJDWLYH WR SRVLWLYH 7KLV UHYHUVHSRODULW\
SURWHFWLRQ HIILFLHQWO\ LQFUHDVHV WKH LVRHOHFWULF SRLQW RI WKH
VHJPHQW DQG LQFUHDVHV LWV VROXELOLW\ ZKLFK OHDGV WR WKH
VXFFHVVIXO V\QWKHVLV RI IXOO\ DFWLYH LQWHUOHXNLQ 7KLV PHWKRG
FDQEHJHQHUDOO\XVHGIRUWKHV\QWKHVLVRIK\GURSKRELF JO\FR
SURWHLQVDQGIXUWKHUVWXGLHVZLOOEHPDGHDORQJWKLVOLQH

◦ 10 ◦◦◦ Division of Protein Chemistry

6ROXELOL]DWLRQRISHSWLGHVHJPHQWE\WKHUHYHUVHSRODULW\SURWHFWLRQDQGLWVDSSOLFDWLRQWRWKHV\QWKHVLVRIKXPDQLQWHUOHXNLQInstitute for Protein Research Osaka University
㻌

㻌

㻌

㻌

㻌 7KH SRVWWUDQVODWLRQDO PRGLILFDWLRQ RI KLVWRQHV SOD\V DQ
‘LPSRUWDQWUROHLQWKHHSLJHQHWLFUHJXODWLRQRIJHQHH[SUHVVLRQ$㻌
VSHFLILFDOO\PRGLILHG KLVWRQH OLEUDU\ LV H[WUHPHO\ XVHIXO IRU㻌

IXQFWLRQDOVWXGLHVRIWKHVHPROHFXOHV7KHUHIRUHZHGHYHORSHG㻌

PHWKRGVIRUWKHV\QWKHVLVRIPRGLILHGKLVWRQHV,QWKHILJXUHIRU
㻌
DQH[DPSOHDPHWKRGLVVKRZQIRUV\QWKHVL]LQJVXFKPRGLILHG㻌
KLVWRQHVE\ OLJDWLQJ FKHPLFDOO\ SUHSDUHG 1WHUPLQDO SHSWLGHV㻌
FRQWDLQLQJVSHFLILF PRGLILFDWLRQV DQG &WHUPLQDOUHFRPELQDQWSHSWLGHEXLOGLQJEORFNVLQZKLFKDSHSWLGHFRQWDLQLQJD&\V3UR
㻌

HVWHU &3(  VHTXHQFH LV XVHG DV D WKLRHVWHU SUHFXUVRU 7KH㻌UHVXOWLQJSURWHLQVFRXOGWKHQEHXVHGWRSUHSDUHDQXFOHRVRPH

㻌

OLEUDU\FRQWDLQLQJSRVWWUDQVODWLRQDOPRGLILFDWLRQV㻌㻌㻌 *O\FRV\ODWLRQ LV RQH RI WKH PDMRU SRVWWUDQVODWLRQDO‘PRGLILFDWLRQVRI SURWHLQV 5HFHQWO\ LW KDV EHFRPH NQRZQ WKDW㻌JO\FDQFKDLQV RQ SURWHLQV DUH HQJDJHG LQ YDULRXV ELRORJLFDO㻌SURFHVVHVVXFKDVFHOOGLIIHUHQWLDWLRQFHOODWWDFKPHQWDQGYLUDO㻌㻌

LQIHFWLRQ +RZHYHU WKH GHWDLOV RI WKH SURFHVVHV DUH QRW IXOO\㻌XQGHUVWRRG 7KH PDMRU SUREOHP LV WKDW WKH VWUXFWXUH RI WKH㻌
JO\FDQFKDLQ LV KLJKO\ KHWHURJHQHRXV ZKLFK PDNHV WKHUHODWLRQVKLSEHWZHHQVWUXFWXUHDQGIXQFWLRQYDJXH7RRYHUFRPH㻌WKLV㻌 SUREOHP ZH KDYH EHHQ HQJDJHG LQ HVWDEOLVKLQJ D IDFLOHPHWKRG IRU JO\FRSURWHLQ V\QWKHVLV ZKLFK HQDEOHV WR SUHSDUHJO\FRSURWHLQV KDYLQJ KRPRJHQHRXV FDUERK\GUDWH VWUXFWXUHV
7KHPHWKRGKDVEHHQDSSOLHGWRWKHV\QWKHVLVRIJO\FRSURWHLQVVXFKDVVDSRVLQ&LPPXQRJOREXOLQGRPDLQRI7,0DQGKXPDQ

LQWHUOHXNLQDQGIXQFWLRQDOVWXGLHVDUHEHLQJFRQGXFWHG

3XEOLFDWLRQV
㻌
㻌




2QH3RW )RXU6HJPHQW /LJDWLRQ XVLQJ 6HOHQR DQG 7KLRHVWHUV 6\QWKHVLV RI 6XSHUR[LGH 'LVPXWDVH 7DNHL 7
$QGRK77DNDR7+RMR+  $QJ&KHP,QW(Gt㻌

3UHSDUDWLRQ RI 6HOHQRLQVXOLQ DV D /RQJ/DVWLQJ ,QVXOLQ $QDORJXH $UDL . 7DNHL 7 2NXPXUD 0 :DWDQDEH 6

$PDJDL<$VDKLQD<0RURGHU/+RMR+,QDED.,ZDRND0  $QJ&KHP,QW(G 

6WUXFWXUH RI WKH 'QPW 5HDGHU 0RGXOH &RPSOH[HG ZLWK D 8QLTXH 7ZR0RQR8ELTXLWLQ 0DUN RQ +LVWRQH +

5HYHDOVWKH%DVLVIRU'1$0HWK\ODWLRQ0DLQWHQDQFH,VKL\DPD61LVKL\DPD$6DHNL<0RULWVXJX.0RULPRWR

' <DPDJXFKL / $UDL 1 0DWVXPXUD 5 .DZDNDPL 7 0LVKLPD < +RMR + 6KLPDPXUD 6 ,VKLNDZD ) 7DMLPD 6

7DQDND.$UL\RVKL06KLUDNDZD0,NHJXFKL0.LGHUD$6XHWDNH,$ULWD.1DNDQLVKL0  0ROHFXODU&HOOt

&KHPLFDO6\QWKHVLVRI2*O\FRV\ODWHG+XPDQ,QWHUOHXNLQE\WKH5HYHUVH3RODULW\3URWHFWLRQ6WUDWHJ\$VDKLQD

<.RPL\D62KDJL$)XMLPRWR57DPDJDNL+1DNDJDZD.6DWR7$NLUD67DNDR7,VKLL$1DNDKDUD<+RMR+ $QJ&KHP,QW(G 

6\QWKHVLV RI KLVWRQH SURWHLQV E\ &3( OLJDWLRQ XVLQJ D UHFRPELQDQW SHSWLGH DV WKH &WHUPLQDO EXLOGLQJ EORFN&KHPRHQ]\PDWLF6\QWKHVLVRI,PPXQRJOREXOLQ'RPDLQRI7LP&DUU\LQJWKH&RPSOH[7\SH1*O\FDQ8VLQJWKH.DZDNDPL7<RVKLNDZD5)XML\RVKL<0LVKLPD<+RMR+7DMLPD66XHWDNH,  -%LRFKHPt

2QHSRW/LJDWLRQ0HWKRG$VDKLQD<.DPLWRUL67DNDR71LVKL1+RMR+  $QJ&KHP,QW(G

◦◦◦ 11 ◦IPR஁പ࣯Ռ஁പ࣯Ռָ‫ڂݜ‬෨໵஁പ࣯Ռָ‫ڂݜ‬෨໵
෨໵㻌ָ‫ڂݜ‬
㻌/DERUDWRU\ RI &HOO 6\VWHPV㻌/DERUDWRU\RI&HOO6\VWHPV㻌㻌㻌㻌㻌
3URIHVVRU
0DULNR2NDGD3K'
PRNDGD#SURWHLQRVDNDXDFMS $VVLVWDQW3URIHVVRU.D]XQDUL,ZDPRWR3K'
6KLJH\XNL0DJL3K'
6X[LDQJ=KDQJ3K'

7KHDLPVRIRXUODERUDWRU\DUHWRGHILQHWKHJHQHUDOUHJXODWRU\UXOHVLQVLJQDOWUDQVGXFWLRQWUDQVFULSWLRQDOQHWZRUNV LQ FHOO GHWHUPLQDWLRQ SURFHVVHV DQG WR DSSO\ WKLV NQRZOHGJH RI UHJXODWRU\ SULQFLSOHV WR WKHXQGHUVWDQGLQJ DQG WUHDWPHQW RI KXPDQ GLVHDVHV )RU WKLV SXUSRVH ZH SHUIRUP TXDQWLWDWLYH
PHDVXUHPHQWV RI WKH WDUJHW ELRORJLFDO V\VWHP XVLQJ YDULRXV H[SHULPHQWDO PHWKRGV DQG LQWHJUDWH WKHKHWHURJHQHRXVGDWDE\PHDQVRIPDWKHPDWLFDO PRGHOLQJDQGLQIRUPDWLFV 8VLQJWKHVHV\VWHPVELRORJ\DSSURDFKHV ZH XQFRYHUHG VHYHUDO XQLTXH SURSHUWLHV RI QHWZRUN WRSRORJLHV LQ VLJQDOWUDQVFULSWLRQQHWZRUNVLQLPPXQHFHOOGHYHORSPHQWDQGFDQFHU1RZZHDSSO\WKHVHDQDO\VHVWRGLIIHUHQWELRFKHPLFDOQHWZRUNVWRXQGHUVWDQGUHJXODWRU\PHFKDQLVPVRIKXPDQGLVHDVHV 

5HVHDUFK3URJUDPVʥ '\QDPLFVRIWUDQVFULSWLRQIDFWRUVDQGJHQHUHJXODWLRQ
ʥ 0DWKHPDWLFDOPRGHOLQJRIVLJQDOLQJQHWZRUNVIRUFHOOUHJXODWLRQ
ʥ 0XOWL2PLFVDQDO\VLVRIKXPDQGLVHDVHPRGHOV
5HVHDUFK7RSLFV‘,QWUDFHOOXODUVLJQDOLQJSDWKZD\VDUHLPSRUWDQWWRFKDUDFWHUL]HWKH DFWLYDWLRQ RI WUDQVFULSWLRQ IDFWRUV DQG WKHLU G\QDPLFV DUH
RIWHQ DVVRFLDWHG ZLWK WKH GHWHUPLQDWLRQ RI SDUWLFXODU FHOOXODUSKHQRW\SHV 2XU HDUOLHU VWXG\ LQGLFDWHG WKDW GXUDWLRQ RI (5.
DFWLYLW\GHWHUPLQHVVZLWFKOLNHDOORUQRQHDFWLYDWLRQRIWKHF
)RVWUDQVFULSWLRQIDFWRUWKHUHE\LQGXFLQJTXDOLWDWLYHFKDQJHVLQ
WKH FHOO IDWH 1DNDNXNL HW DO &HOO   2XU VWXG\ RQ %&5
PHGLFDWHG1)N%VLJQDOLQJSDWKZD\VKRZHGWKHFKDUDFWHULVWLF
G\QDPLFV RI 1)N% VZLWFKOLNH DFWLYDWLRQ DQG RVFLOODWRU\
EHKDYLRUVDUHVKDSHGE\IHHGEDFNUHJXODWLRQV,QSDUWLFXODUIRU
D PHFKDQLVP RI VZLWFKOLNH DFWLYDWLRQ ZH IRXQG WKDW ,..
DFWLYLW\ LV UHJXODWHG E\ SRVLWLYH IHHGEDFN IURP ,.. WR 7$.
PHGLDWHGE\VFDIIROGLQJSURWHLQ&$50$DW6HUDQGWKLV
IHHGEDFNUHJXODWLRQFRXOGLQGXFHWKHVZLWFKOLNHDFWLYDWLRQRI1)N% LQ VLQJOH FHOOV 6KLQRKDUD HW DO 6FLHQFH   2XU
VWXGLHV LQGLFDWH WKDW WRSRORJLHV RI WKH VLJQDOLQJ QHWZRUN DUH
LPSRUWDQWIRUHIILFLHQWVLJQDOWUDQVPLVVLRQLQWKHFHOO

◦ 12 ◦◦◦ Division of Protein Chemistry)LJ$OORUQRQHDFWLYDWLRQRI1)N%WUDQVFULSWLRQIDFWRU LV GHWHUPLQHG E\ SRVLWLYH IHHGEDFNUHJXODWLRQVLQWKHVLJQDOLQJQHWZRUN

Institute for Protein Research Osaka University‘ '\VUHJXODWHG G\QDPLFV RI VLJQDOLQJ UHVSRQVHV DUH FORVHO\㻌UHODWHGWRLQFLGHQFHRIVHYHUDOW\SHVRIKXPDQFDQFHUWKHUHIRUH

㻌 TXDQWLWDWLYH XQGHUVWDQGLQJ RI FDQFHU QHWZRUN LV LPSRUWDQW
WKH
㻌 WKH EHWWHU XQGHUVWDQGLQJ RI GLVHDVH PHFKDQLVPV DQG
IRU
GHYHORSPHQWRIHIIHFWLYHWUHDWPHQWV2XUODERUDWRU\GHYHORSHG㻌DPDWKHPDWLFDOPRGHORIYDULRXVVLJQDOLQJSDWKZD\VEDVHGRQ㻌KLJKO\TXDQWLWDWLYHH[SHULPHQWDOGDWD%DVHGRQWKLVNQRZOHGJH㻌ZHDUHQRZGHYHORSLQJDZKROHFHOOVLJQDOLQJQHWZRUNRIKXPDQ㻌
FDQFHU:H EHOLHYH WKDW WKH PRGHO ZLOO EH XWLOL]HG IRU GUXJ㻌GLVFRYHU\FHOOVSHFLILF GUXJ GHYHORSPHQW DQG LGHQWLILFDWLRQ RI

GUXJUHVLVWDQWPDUNHUV㻌)LJ  0DWKHPDWLFDO PRGHOLQJ RI WKH ZKROH FHOOVLJQDOLQJQHWZRUN ‘$GDSWRU SURWHLQV DUH PROHFXODU KXEV RI VLJQDOLQJ QHWZRUNV㻌DQGPHGLDWHDYDULHW\RIVLJQDOLQJSDWKZD\VZKLFKDUHHVVHQWLDO㻌LQFHOOXODUVXUYLYDODQGGHYHORSPHQWDQGLWLPSOLHVWKDWPXOWLRPLFV
DQDO\VLV㻌㻌 RI DGDSWRU SURWHLQ NQRFNRXW FHOOV PD\ UHYHDO D
IXQGDPHQWDO
UROH RI WKH VLJQDOLQJ SDWKZD\ LQ RYHUDOO FHOO㻌IXQFWLRQV :H SHUIRUP WUDQVFULSWRPH HSLJHQHWLFV DQG

PHWDERORPH DQDO\VLV WRJHWKHU ZLWKVHYHUDO W\SHV RI SURWHRPLF㻌

DQDO\VHVRIFHOOVRULJLQDWHGIURPDGDSWRUSURWHLQNQRFNRXWPLFH㻌7KH
DQDO\VLV ZLOO SURYLGH XV WKH EHWWHU XQGHUVWDQGLQJ RI WKH
EDVLF
SULQFLSOHV RI KRZ VLJQDOLQJ SDWKZD\V OLQN WR RWKHU㻌ELRFKHPLFDOQHWZRUNV WR GHWHUPLQH WKH SKHQRW\SLF RXWSXWV RI
㻌WKHFHOOVDQGLQGLYLGXDOV)LJ$GDSWRUSURWHLQNQRFNRXWLQGXFHVGUDPDWLF
FKDQJHVLQWUDQVFULSWRPHODQGVFDSH

3XEOLFDWLRQV㻌 7UDQVFULSWLRQDOO\LQGXFLEOH3+/'$DWWHQXDWHVWKHDFWLYLW\RI(UE%UHFHSWRUV0DJL6HWDO  -%LRO
&KHPGRLMEF0&XUUHQW6WDWXVRI0DWKHPDWLFDO0RGHOLQJRI&DQFHUt)URPWKH9LHZSRLQWRI&DQFHU+DOOPDUNV0DJL6HWDO

2VFLOODWLRQG\QDPLFVXQGHUOLHVIXQFWLRQDOVZLWFKLQJRI1)Ж%IRU%FHOODFWLYDWLRQ,QRXH.HWDO  QSM &XUUHQW2SLQLRQLQ6\VWHPV%LRORJ\GRLMFRLVE

6\VWHPV%LRORJ\ $SSOLFDWLRQVGRLQSMVED

3RVLWLYHIHHGEDFNZLWKLQDNLQDVHVLJQDOLQJFRPSOH[IXQFWLRQVDVDVZLWFKPHFKDQLVPIRU1)Ж%DFWLYDWLRQ
6KLQRKDUD+HWDO  6FLHQFHGRLVFLHQFH,QWHJUDWHGTXDQWLWDWLYHDQDO\VLVRIWKHSKRVSKRSURWHRPHDQGWUDQVFULSWRPHLQWDPR[LIHQUHVLVWDQWEUHDVW
FDQFHU2\DPD0HWDO  -%LRO&KHPGRLMEF0/LJDQGVSHFLILFF)RVH[SUHVVLRQHPHUJHVIURPWKHVSDWLRWHPSRUDOFRQWURORI(UE%QHWZRUNG\QDPLFV/LJDQGGHSHQGHQWUHVSRQVHVRIWKH(UE%VLJQDOLQJQHWZRUNH[SHULPHQWDODQGPRGHOLQJDQDO\VLV%LUWZLVWOH05



1DNDNXNL7HWDO  &HOOGRLMFHOO
HWDO  0RO6\VW%LROGRLKWWSPVE

4XDQWLWDWLYHWUDQVFULSWLRQDOFRQWURORI(UE%UHFHSWRUVLJQDOLQJXQGHUJRHVJUDGHGWRELSKDVLFUHVSRQVHIRUFHOOGLIIHUHQWLDWLRQ1DJDVKLPD7HWDO  -%LRO&KHPGRLMEF0

◦◦◦ 13 ◦IPR஁പ࣯ߑ஁പ࣯ߑଆਫ਼෼ָ‫ڂݜ‬෨໵஁പ࣯ߑଆਫ਼෼ָ
෼ָ‫ڂݜ‬෨෨໵㻌㻌

/DERUDWRU\RI1DQRELRORJ\
㻌/DERUDWRU\RI1DQRELRORJ\㻌
3URIHVVRU<RVKLH+DUDGD3K'\KDUDGD#SURWHLQRVDNDXDFMS $VVRFLDWH3URIHVVRU0DGRND6X]XNL3K'.

$VVLVWDQW3URIHVVRU+LVDVKL7DGDNXPD3K'%LRPROHFXOHVDUHFRQVWDQWO\SHUWXUEHGE\WKHVXUURXQGLQJHQYLURQPHQW7KXVLQFRQWUDVWWRWKHDUWLILFLDOV\VWHPELRPROHFXOHVKDUQHVVWKHUPDOIOXFWXDWLRQDFKLHYLQJKLJKHQHUJ\HIILFLHQF\)RUH[DPSOHZLWKRXWXVLQJFKHPLFDOHQHUJ\VXFKDV$7351$SRO\PHUDVHVGLIIXVHRQHGLPHQVLRQDOO\RQ'1$WRVHDUFKWKH

SURPRWHU VHTXHQFH 2XU XOWLPDWH JRDO LV WR HOXFLGDWH WKH VRSKLVWLFDWHG PROHFXODU PHFKDQLVP RI

ELRPROHFXOHV,PDJLQJ DQG PDQLSXODWLQJ LQGLYLGXDO PROHFXOHV DUH SRZHUIXO WHFKQLTXHV :H KDYH GHYHORSHG QRYHOWHFKQRORJ\VXFKDVVLQJOHPROHFXOHLPDJLQJDQGKDYHYLVXDOL]HGWKHPRWLRQDQGRUVWUXFWXUDOFKDQJHRIELRPROHFXOHVDWWKHVLQJOHPROHFXOHOHYHOUHYHDOLQJVRPHVHFUHWV RIWKHQDWXUH2XU FXUUHQWWRSLFVDUH

GHYHORSLQJQRYHOWRROVWRLQYHVWLJDWHWKHWKHUPDOELRORJ\DQGJHQHWLFV

5HVHDUFK3URMHFWV'HYHORSLQJQRYHOVLQJOHPROHFXOHLPDJLQJWHFKQLTXHVXVLQJIOXRUHVFHQWGLDPRQG'HYHORSLQJWHFKQLTXHVIRUORFDOWHPSHUDWXUHPHDVXUHPHQWLQYLYR$QDO\VLVRIELRPROHFXODULQWHUDFWLRQVZLWK]HURPRGHZDYHJXLGHV =0:V



$QDO\VLVRIWUDQVFULSWLRQDOUHJXODWLRQXVLQJWKH'1$RULJDPL

5HVHDUFK7RSLFV 1DQRGLDPRQGV&RQYHQWLRQDOIOXRUHVFHQWSUREHVKDYHLQWULQVLFSUREOHPVVXFKDV SKRWREOHDFKLQJEOLQNLQJ KLQGHUDQFHE\DXWRIOXRUHVFHQFH RI WKH ELRORJLFDO VDPSOH   ORZ UHVROXWLRQ RI DQJOH YDULDWLRQ DQG URWDWLRQDO PRWLRQ 7R UHVROYH VXFKSUREOHPVZHKDYHGHYHORSHGIOXRUHVFHQWQDQRGLDPRQGV )1'V 8SRQQPYLVLEOHOLJKWH[FLWDWLRQWKH1LWURJHQ
9DFDQF\ &HQWHU 19&  RI WKH GLDPRQG HPLWV D IOXRUHVFHQW VLJQDO ,Q FRQWUDVW WR FRQYHQWLRQDO IOXRUHVFHQW SUREHVIOXRUHVFHQFHIURP19&LVQRWSKRWREOHDFKHGRUEOLQNHGDOORZLQJXVSURORQJHGREVHUYDWLRQRIELRORJLFDOVDPSOHV)XUWKHUPRUHXVLQJDQRSWLFDOO\GHWHFWHGPDJQHWLFUHVRQDQFH 2'05 WHFKQLTXHZHKDYHVXFFHHGLQGLVWLQJXLVKLQJWKH )1'V IOXRUHVFHQW VLJQDO IURP WKDW RI DXWRIOXRUHVFHQFH LQ WKH &HOHJDQV :H DUH FXUUHQWO\ GHYHORSLQJ KLJKO\VHQVLWLYHPHDVXUHPHQWPHWKRGVRIURWDWLRQDOPRWLRQ㻌 㻌 㻌 㻌 &RQFHSWRIIOXRUHVFHQWQDQRGLDPRQGV◦ 14 ◦◦◦ Division of Protein Chemistry  )OXRUHVFHQWLPDJHRI)1'V

Institute for Protein Research Osaka University㻌஁പ࣯ߑ஁പ࣯ߑଆਫ਼෼ָ‫ڂݜ‬෨໵
஁പ
࣯ߑଆਫ਼
ଆਫ਼෼ָ
෼ָ‫ڂݜ‬
෼ָ‫ڂݜ‬෨໵
‫ڂݜ‬෨໵ଆਫ਼ 7KHUPDOELRORJ\
7HPSHUDWXUH LV WKH IXQGDPHQWDO SDUDPHWHU RI ELRORJ\ DIIHFWLQJ PHWDEROLVP DQG ELRUK\WKP /RFDO WHPSHUDWXUHIOXFWXDWLRQ LQVLGH WKHLQGLYLGXDO FHOOLV HQLJPDWLF :H KDYH GHYHORSHG D WKHUPRPHWHU XVLQJ KLJKPROHFXODUZHLJKWSRO\PHUDQG)1'V:LWKTXDQWLWDWLYHPHDVXUHPHQWVZHWDFNOHWKHELRORJLFDOUHOHYDQFHRIWKHWKHUPDOIOXFWXDWLRQ
7KURXJKSURRILQJRIWKHFRQFHSWRI7HPSHUDWXUHVLJQDOLQJZHDLPWRHVWDEOLVKWKH7KHUPDOELRORJ\
㻌

㻌
㻌
㻌
㻌
㻌
/LQHDUSRO\PHULFWKHUPRPHWHU      ,PDJLQJRILQWUDFHOOXODUWHPSHUDWXUH㻌 (SLJHQHWLFLPDJLQJ㻌
7KHPD[LPXPIOXRUHVFHQWVLJQDOFRQFHQWUDWLRQRIFRQYHQWLRQDOVLQJOHPROHFXOHWHFKQLTXHVVXFKDV7,5)PLFURVFRS\
LVDERXWQ0OLPLWLQJWKHYHUVDWLOLW\RIWKHLUDSSOLFDWLRQ7RRYHUFRPHWKLVLVVXHZHKDYHGHYHORSHGQHZWHFKQLTXHVXVLQJ ]HURPRGH ZDYHJXLGHV =0:V  ZKLFK DOORZ XV WR REVHUYH VLQJOH PROHFXOH LQWHUDFWLRQ XQGHU PLFUR0
IOXRUHVFHQWVLJQDOV:HDSSOLHG=0:VWRHOXFLGDWHWKHPROHFXODUPHFKDQLVPRIJHQHH[SUHVVLRQFRQWURO







6FKHPDWLFLOOXVWUDWLRQRI=0:V                     6(0DQGIOXRUHVFHQWLPDJHRI=0:V

Publications










(QULFKPHQWRI2'05DFWLYHQLWURJHQYDFDQF\FHQWUHVLQILYHQDQRPHWUHVL]HGGHWRQDWLRQV\QWKHVL]HG

QDQRGLDPRQGV1DQRSUREHVIRUWHPSHUDWXUHDQJOHDQGSRVLWLRQ6RWRPD6HWDO6FL5HS  
 

&DDVVRFLDWHGWULSKDVLFS+FKDQJHVLQPLWRFKRQGULDGXULQJEURZQDGLSRF\WHDFWLYDWLRQ+RX<HWDO0RO

0HWDEROLVP    *ROG1DQRVKHOO0HGLDWHG5HPRWH0\RWXEH$FWLYDWLRQ0DULQR$HWDO$&61DQR    $%HHWOH)OLJKW0XVFOH'LVSOD\V/HJ0XVFOH0LFURVWUXFWXUH6KLPRPXUD7HWDO%LRSK\VLFDOMRXUQDO     

6LQJOH0ROHFXOH$QDO\VLVRIWKH7DUJHW&OHDYDJH5HDFWLRQE\WKH'URVRSKLOD51$L(Q]\PH&RPSOH[<DR&HW
DO0RO&HOO  

6\QHUJLVWLFHIIHFWRI$73IRU5XY$5XY%+ROOLGD\MXQFWLRQ'1$FRPSOH[IRUPDWLRQ,ZDVD7HWDO6FL5HS

  



5HDOWLPHEDFNJURXQGIUHHVHOHFWLYHLPDJLQJRIIOXRUHVFHQWQDQRGLDPRQGVLQYLYR,JDUDVKL5HWDO1DQR/HWW,QWUDFHOOXODUWHPSHUDWXUHPDSSLQJZLWKDIOXRUHVFHQWSRO\PHULFWKHUPRPHWHUDQGIOXRUHVFHQFHOLIHWLPHLPDJLQJ  

PLFURVFRS\2NDEH.HWDO1DW&RPPXQ  ◦◦◦ 15 ◦IPR㻌/DERUDWRU\ RI 2UJDQHOOH %LRORJ\
㻌/DERUDWRU\RI2UJDQHOOH%LRORJ\㻌㻌㻌,QGHSHQGHQW$VVRFLDWH3URIHVVRU

0DVDWR1DNDL'6FL
QDNDL#SURWHLQRVDNDXDFMS +LJKHU SODQWV PXVW LPSRUW f GLIIHUHQW F\WRVROLFDOO\V\QWKHVL]HGQXFOHDUHQFRGHGFKORURSODVWSURWHLQVDFURVV
WKH GRXEOH HQYHORSH PHPEUDQHV VXUURXQGLQJ WKHRUJDQHOOH WR IXOILOO WKHLU FRPSOH[ SK\VLRORJLFDO UROHVLQFOXGLQJ SKRWRV\QWKHWLF IXQFWLRQV 7ZR VXFFHVVLYH
SURWHLQ WUDQVORFRQV DW WKH RXWHU DQG LQQHU HQYHORSH
PHPEUDQHV WHUPHG 72& DQG 7,& UHVSHFWLYHO\ DUHUHVSRQVLEOHIRUSURWHLQLPSRUWLQWRFKORURSODVWV2XUUHFHQWGLVFRYHU\ RI WKH JHQXLQH 7,& WUDQVORFRQ SXEOLVKHG LQ
6FLHQFH  LQ    SURYLGHV DQ HQWLUHO\ UHYLVHG YLHZ RQ
WKHPROHFXODUPHFKDQLVPVRISURWHLQWUDQVORFDWLRQDFURVVWKHLQQHUHQYHORSHPHPEUDQHRIFKORURSODVW  

5HVHDUFK3URJUDPV(OXFLGDWLRQRIPROHFXODUPHFKDQLVPVRISURWHLQLPSRUWLQWRWKHFKORURSODVWV
0ROHFXODUHYROXWLRQDU\VWXGLHVRQWKHFKORURSODVWSURWHLQLPSRUWV\VWHP

3XEOLFDWLRQV
 8QFRYHULQJ WKH 3URWHLQ 7UDQVORFRQ DW WKH &KORURSODVW ,QQHU (QYHORSH 0HPEUDQH  6 .LNXFKL 1DNDL 0 HW DO 6FLHQFH㻌 
 1HZSHUVSHFWLYHVRQ&KORURSODVW3URWHLQ,PSRUW1DNDL0  3ODQW&HOO3K\VLRO 7KH7,&FRPSOH[XQFRYHUHG7KHDOWHUQDWLYHYLHZRQWKHPROHFXODUPHFKDQLVPRISURWHLQWUDQVORFDWLRQDFURVVWKHLQQHUHQYHORSHPHPEUDQHRIFKORURSODVWV1DNDL0  %LRFKLP%LRSK\V$FWD
 <&)$*UHHQ7,&1DNDL0  3ODQW&HOO

◦ 16 ◦◦◦ Division of Protein Chemistry

Division of Protein
Structural Biology

Laboratory of Protein Folding
Laboratory of Molecular Biophysics
Laboratory of Protein Crystallography
Laboratory of Membrane Protein Chemistry

IPR㻌/DERUDWRU\ RI 3URWHLQ )ROGLQJ
㻌/DERUDWRU\RI3URWHLQ)ROGLQJ㻌㻌㻌
3URIHVVRU<XML*RWR'6FL
JW\M#SURWHLQRVDNDXDFMS 

6SHFLDOO\DSSRLQWHG/HFWXUHU.HQML6DVDKDUD'6FL$VVLVWDQW3URIHVVRU 

0DVDWRPR6R'6FL

6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU.HLLFKL<DPDJXFKL'6FL

3URWHLQIROGLQJLVDSURFHVVLQZKLFKDQH[WHQGHGSRO\SHSWLGHFKDLQDFTXLUHVDXQLTXHIROGHGFRQIRUPDWLRQZLWK ELRORJLFDO DFWLYLW\ &ODULI\LQJ WKH PHFKDQLVP RI SURWHLQ IROGLQJ LV HVVHQWLDO WR LPSURYH RXUXQGHUVWDQGLQJ RI WKH VWUXFWXUH DQG IXQFWLRQ RI SURWHLQV ,W LV DOVR LPSRUWDQW EHFDXVH PDQ\ FULWLFDOELRORJLFDOSURFHVVHVDQGGLVHDVHVWDWHVLQYROYHSURWHLQPLVIROGLQJDQGDJJUHJDWLRQUHDFWLRQV:HVWXG\

WKH FRQIRUPDWLRQDO VWDELOLW\ DQG WKH PHFKDQLVPV RI SURWHLQ IROGLQJ DQG PLVIROGLQJ ZLWK YDULRXVDSSURDFKHV LQFOXGLQJ VSHFWURVFRSLHV 105 IOXRUHVFHQFH &'  SK\VLFRFKHPLFDO PHDVXUHPHQWVFDORULPHWU\DQDO\WLFDOXOWUDFHQWULIXJDWLRQ DQGIOXRUHVFHQFHPLFURVFRS\

5HVHDUFK3URJUDPV)ROGLQJDQGVWDELOLW\RISURWHLQV

0HFKDQLVPRIIRUPDWLRQRIDP\ORLGILEULOV&RPSUHKHQVLYHXQGHUVWDQGLQJRISURWHLQIROGLQJDQGPLVIROGLQJ

5HVHDUFK7RSLFV‘ +HDWRIDP\ORLGEXUVWGLUHFWO\PRQLWRUHGE\LVRWKHUPDOWLWUDWLRQFDORULPHWU\$P\ORLG ILEULOV IRUP LQ VXSHUVDWXUDWHG VROXWLRQV YLD D QXFOHDWLRQ DQG JURZWK PHFKDQLVP :LWK ЎPLFURJOREXOLQЎP DSURWHLQUHVSRQVLEOHIRUGLDO\VLVUHODWHGDP\ORLGRVLVZHVKRZHGGLUHFWKHDWPHDVXUHPHQWVRIWKHIRUPDWLRQ
RI DP\ORLG ILEULOV XVLQJ LVRWKHUPDO WLWUDWLRQ FDORULPHWU\ ,7&    ,7& ZLOO EHFRPH D SURPLVLQJ DSSURDFK IRU
FRPSUHKHQVLYHO\FODULI\LQJWKHWKHUPRG\QDPLFVRISURWHLQIROGLQJDQGPLVIROGLQJ

㻌)LJ  /HIW  )LEULOODWLRQ RI EPLFURJOREXOLQ PRQLWRUHG E\ ,7& DQG 5LJKW  VLPLODULW\ RI DP\ORLG
ILEULOODWLRQDQGFU\VWDOOL]DWLRQRIVRGLXPDFHWDWH

◦ 18 ◦◦◦ Division of Protein Structural Biology

Institute for Protein Research Osaka University஁പ࣯ߑଆਫ਼෼ָ‫ڂݜ‬෨໵
஁
஁പ
പ࣯ߑ
࣯ߑଆਫ਼
ଆਫ਼෼ָ
෼ָ‫ڂݜ‬
෼ָ‫ڂݜ‬෨໵
‫ڂݜ‬෨໵㻌ଆਫ਼
‘ 8OWUDVRQLFDWLRQDFFHOHUDWHGDP\ORLGILEULOODWLRQ$P\ORLGILEULOVIRUPLQVXSHUVDWXUDWHGVROXWLRQVYLDDQXFOHDWLRQDQGJURZWKPHFKDQLVP:HIRXQGWKDWXOWUDVRQLFDWLRQHIIHFWLYHO\EUHDNVVXSHUVDWXUDWLRQDQGIRUFHVILEULOODWLRQ%\FRQVWUXFWLQJD+$1GDL$P\ORLG%XUVW,QGXFHU +$1$%, 
ZKLFKFRPELQHVWKHXVHRIDZDWHUEDWKW\SHXOWUDVRQLFDWRUDQGDPLFURSODWHUHDGHUZHVWXG\WKHXOWUDVRQLFDWLRQIRUFHGILEULOODWLRQRIVHYHUDOSURWHLQVZLWKDIRFXVRQIOXFWXDWLRQLQWKHODJWLPH

)LJ /HIW XOWUDVRQLFDWLRQIRUFHGDP\ORLGILEULOODWLRQPRQLWRUHG
E\7K7DQG 5LJKW VFKHPDWLFLOOXVWUDWLRQVRID+$1$%,V\VWHP

‘ &RPSUHKHQVLYHXQGHUVWDQGLQJRISURWHLQDJJUHJDWLRQRQWKHEDVLVRIVROXELOLW\DQGVXSHUVDWXUDWLRQ$P\ORLGILEULOVDUHIRUPHGE\DQXFOHDWLRQJURZWKPHFKDQLVPVLPLODUWRWKHFU\VWDOOL]DWLRQRIVROXWHVIURPVROXWLRQ
6ROXELOLW\DQGVXSHUVDWXUDWLRQDUHWZRRIWKHPRVWLPSRUWDQWIDFWRUVGHWHUPLQLQJFU\VWDOOL]DWLRQRIVROXWHV0RUHRYHU
FU\VWDOOL]DWLRQFRPSHWHVZLWKJODVVIRUPDWLRQLQZKLFKVROXWHVFROODSVHLQWRDPRUSKRXVDJJUHJDWHV8QGHUVWDQGLQJ
WKH UROH RI VXSHUVDWXUDWLRQ LQ GHWHUPLQLQJ DJJUHJDWLRQEDVHG SKDVHWUDQVLWLRQV RI GHQDWXUHG SURWHLQV SURYLGHV DQ
LPSRUWDQWFRPSOHPHQWDU\SRLQWRIYLHZWRVWUXFWXUDOVWXGLHVRISURWHLQDJJUHJDWHV  )LJ /HIW )UHHHQHUJ\SURILOHVIRUWKHFRPSHWLWLYHIRUPDWLRQRIDPRUSKRXVDJJUHJDWHVDQGDP\ORLGILEULOV 0LGGOH VLPLODULW\RIWKHSKDVHGLDJUDPVIRUDP\ORLGILEULOODWLRQDQGSURWHLQFU\VWDOOL]DWLRQDQG 5LJKW DQHZYLHZRIDP\ORLGILEULOODWLRQRI SURWHLQV

3XEOLFDWLRQV






+HSDULQGHSHQGHQWDJJUHJDWLRQRIKHQHJJZKLWHO\VR]\PHUHYHDOVWZRGLVWLQFWPHFKDQLVPVRIDP\ORLG

ILEULOODWLRQ1LWDQL$HWDO  -%LRO&KHP

5HYLVLWLQJVXSHUVDWXUDWLRQDVDIDFWRUGHWHUPLQLQJDP\ORLGILEULOODWLRQ6R0+DOO'*RWR<  &XUU2SLQ

6WUXFW%LRO

6XSHUVDWXUDWLRQOLPLWHGDQGXQOLPLWHGSKDVHWUDQVLWLRQVFRPSHWHWRSURGXFHWKHSDWKZD\FRPSOH[LW\LQ

DP\ORLGILEULOODWLRQ$GDFKL06R06DNXUDL..DUGRV-*RWR<  -%LRO&KHP 

+HDWRIVXSHUVDWXUDWLRQOLPLWHGDP\ORLGEXUVWGLUHFWO\PRQLWRUHGE\LVRWKHUPDOWLWUDWLRQFDORULPHWU\,NHQRXH7

/HH<+.DUGRV-<DJL+,NHJDPL71DLNL+*RWR<  3URF1DWO$FDG6FL86$

◦◦◦ 19 ◦IPR஁പ࣯ߑଆਫ਼෼ָ‫ڂݜ‬෨໵
஁
஁പ
പ࣯ߑ
࣯ߑଆਫ਼
ଆਫ਼෼ָ
෼ָ‫ڂݜ‬
෼ָ‫ڂݜ‬෨໵
‫ڂݜ‬
෨໵
㻌ଆਫ਼
㻌

/DERUDWRU\ RI 0ROHFXODU %LRSK\VLFV㻌/DERUDWRU\RI0ROHFXODU%LRSK\VLFV㻌㻌㻌
3URIHVVRU7RVKLPLFKL)XMLZDUD'6FLWIMZU#SURWHLQRVDNDXDFMS $VVRFLDWH3URIHVVRU

<RK0DWVXNL3K'

6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU  $\DNR(JDZD3K'

7RVKLKLNR6XJLNL3K'  .HQLFKL+DUDGD3K'-XQ)XND]DZD3K'.\RNR)XUXLWD3K' 

7KH/DERUDWRU\RI0ROHFXODU%LRSK\VLFVLVHQJDJHGLQVWXG\LQJWKHELRORJLFDOPDFURPROHFXODUVWUXFWXUH

DQG LWV LQWHUPROHFXODU LQWHUDFWLRQV PDLQO\ E\ XVLQJ QXFOHDU PDJQHWLF UHVRQDQFH 105  105 SURYLGHVLQIRUPDWLRQRQWKHSURWHLQVWUXFWXUHDWZRUNZLWKDWRPLFUHVROXWLRQHYHQLQFHOOV7KLVDGYDQWDJHDOORZVXVWRXQGHUVWDQGWKHELRORJLFDODFWLYLWLHVIRUVLJQDOWUDQVGXFWLRQDQGHQHUJ\FRQYHUVLRQIURPVWUXFWXUHV6WUXFWXUHV RI )R FULQJ OLJKWKDUYHVWLQJ %FKO F FRPSOH[ E PLFURJOREXOLQ DP\ORLG IORULJHQ LQWHUDFWLQJXELTXLWLQ DQG PHPEUDQHERXQG PDVWRSDUDQ; KDYH EHHQ HOXFLGDWHG 6LQFH VXSUDPROHFXODU V\VWHPVVXFKDVPHPEUDQHSURWHLQFRPSOH[HVSOD\LPSRUWDQWUROHVLQELRORJLFDOV\VWHPVZHDUHDOVRGHYHORSLQJQHZPHWKRGRORJLHVLQ105WRDQDO\]HWKRVHFKDOOHQJLQJVWUXFWXUHV2QHRIRXUSURJUDPVIRUVROLGVWDWH

105IHDWXUHVKLJKILHOGG\QDPLFQXFOHDUSRODUL]DWLRQ '13 IRUDIROGVLJQDOHQKDQFHPHQWE\XVLQJKLJKLQWHQVLW\WHUDKHUW]OLJKWVRXUFHJ\URWURQ7KHVHGHYHORSPHQWVDLPWRFRQWULEXWHWRQRWRQO\DFDGHPLF
EXWDOVRLQGXVWULDO105DSSOLFDWLRQVVXFKDVGUXJGLVFRYHU\

5HVHDUFK3URJUDPV105DQDO\VLVRISURWHLQVDQGWKHLULQWHUDFWLRQV

1HZ PHWKRGRORJLHV LQ ELRORJLFDO 105 LQFOXGLQJ LVRWRSHODEHOHG VDPSOH SUHSDUDWLRQ DQG GDWD



6HQVLWLYLW\HQKDQFHPHQWRIKLJKUHVROXWLRQ105E\K\SHUSRODUL]DWLRQ

DQDO\VLV

5HVHDUFK7RSLFV+LJKUHVROXWLRQ 105 DQDO\VLV RI VWUXFWXUDO DQG IXQFWLRQDO RI SURWHLQV LQ VROXWLRQV DQG ELRPHPEUDQHV UHTXLUHVGHYHORSPHQWRIPHWKRGVIRUVDPSOHSUHSDUDWLRQ105H[SHULPHQWVDQGGDWDDQDO\VLV   105 VSHFWURVFRS\ FDQ SURYLGH WKH GHWDLOHG LQIRUPDWLRQ HYHQ RQ WUDQVLHQW SURWHLQOLJDQG
DQG SURWHLQSURWHLQ㻌LQWHUDFWLRQV 7R SHUIRUP VXFK DQDO\VLV IRU FKDOOHQJLQJ WDUJHWV ZLWK ODUJH PROHFXODU PDVVHV DQG ORZ \LHOGV ZH
LQWURGXFHGPHWKRGRORJ\IRUVDPSOHSUHSDUDWLRQVVXFKDVSRVWWUDQVODWLRQDOSURWHLQPRGLILFDWLRQZLWKKLJKVHQVLWLYLW\
PROHFXODUSUREHVHIILFLHQWDPLQRDFLGVHOHFWLYHLVRWRSHODEHOLQJDQG)DGGLWLRQWROLJDQGPROHFXOHVDVDVHQVLWLYH
VSLQ
)LJ   +t& +64& VSHFWUD RI PHWK\ODWHG
XELTXLWLQDQGPHWK\ODWHGXELTXLWLQLQWHUDFWLQJZLWKSURWHLQ <8+ VKRZQ RQ WKH OHIW 8ELTXLWLQ DQG<8+ DUH VKRZQ LQ WKH FRPSOH[ &0;  E\ WKHULEERQ DQG VXUIDFH UHSUHVHQWDWLRQ UHVSHFWLYHO\㻌/DUJHU DQG VPDOOHU FKHPLFDO VKLIW FKDQJHV DUHFRORUHG UHG DQG JUD\ UHVSHFWLYHO\ 7KLV VLPSOH
SRVWPHWK\ODWLRQ PHWKRG JLYHV VWURQJ &+105
VLJQDOV IRU GHWHFWLQJ WKH SURWHLQ LQWHUDFWLRQV
5HI ◦ 20 ◦◦◦ Division of Protein Structural Biology

Institute for Protein Research Osaka University㻌஁പ࣯ߑ஁പ࣯ߑଆਫ਼෼ָ‫ڂݜ‬෨໵
஁പ
࣯ߑଆਫ਼
ଆਫ਼෼ָ
෼ָ‫ڂݜ‬
෼ָ‫ڂݜ‬෨໵‫ڂݜ‬෨໵㻌ଆਫ਼㻌  7KHPDJQHWLF LQWHUDFWLRQ RI HOHFWURQ VSLQV LV DERXW  WLPHV VWURQJHU WKDQ WKDW RI QXFOHDU VSLQV :H DUH
㻌GHYHORSLQJKLJKILHOG'13105VSHFWURPHWHUVIRUIROG105VHQVLWLYLW\HQKDQFHPHQWXVLQJHOHFWURQVSLQVDW㻌FU\RJHQLF WHPSHUDWXUHV LQ FROODERUDWLRQ ZLWK VFLHQWLVWV DQG HQJLQHHUV LQ DFDGHPLDDQG LQGXVWULHV 7KLV XOWUDKLJKVHQVLWLYLW\VSHFWURPHWHUVDUHDYDLODEOHWKURXJKVKDUHGXVHSURJUDPVDW,35

㻌
㻌
㻌
㻌
㻌

)LJ   105 PDJQHW IRU 0+] VROLGVWDWH 105 ZLWK D
FORVHGF\FOHFROGKHOLXPPDJLFDQJOHVSLQQLQJV\VWHPRQWKH
OHIW DQG *+] J\URWURQ IRU KLJKLQWHQVLW\ OLJKW VRXUFH RI
WHUDKHUW]ZDYHRQWKHULJKW+\SHUSRODUL]DWLRQJHQHUDWHGZLWKWKHVHLQVWUXPHQWV LQFUHDVHV WKH 105 VHQVLWLYLW\ RISURWHLQV
7KLV '13105 VSHFWURPHWHU ZDV GHYHORSHG LQ WKH ,QVWLWXWH
IRU3URWHLQ5HVHDUFK 5HI 

㻌
㻌
㻌
㻌
㻌
㻌
㻌
㻌

 :HFDQQRZILQGUHODWLRQVKLSVEHWZHHQ105SDUDPHWHUVDQGSURWHLQVWUXFWXUHVEDVHGRQDKXJHQXPEHURISURWHLQ㻌VWUXFWXUHV LQ GDWDEDVH 3'% DQG XVLQJ FRPSXWDWLRQDO FKHPLVWU\ TXDQWXP PHFKDQLFDO DQG PROHFXODU G\QDPLFVFDOFXODWLRQV  %\PDNLQJXVHRIVXFKUHODWLRQVKLSVZHHIILFLHQWO\FRQGXFWVWUXFWXUDODQGIXQFWLRQDODQDO\VLVRISURWHLQV\VWHPV)LJ  $XWRPDWLFVROLGVWDWH105VWUXFWXUDODQDO\VLVRI
SURWHLQ&105VSHFWUDRISURWHLQVLQVROLGVRIWHQVKRZXQUHVROYHGVLJQDOV2XUVSHFWUDOILWWLQJVRIWZDUH5(63/6
HQDEOHV FKHPLFDO VKLIW DVVLJQPHQWV DQG VHFRQGDU\VWUXFWXUH SUHGLFWLRQ EDVHG RQ WKH GDWDEDVHV 3'%M DQG%05%7KLVPHWKRGVLPSOLILHVWKHVWUXFWXUDODQDO\VLVE\SURYLGLQJUHOLDEOHLQIRUPDWLRQHYHQIRUO\RSKLOL]HGVWDWHV
5HI 
3XEOLFDWLRQV㻌0HFKDQLVWLF DQG VWUXFWXUDO EDVLV RI ELRHQJLQHHUHG ERYLQH &DWKHOLFLGLQ ZLWK RSWLPL]HG WKHUDSHXWLF DFWLYLW\㻌

6DKRR%0DUX\DPD.(GXOD-7RXJDQ7/LQ</HH<++RULL7DQG)XMLZDUD7   6FL5HS

6WUXFWXUHGHWHUPLQDWLRQRIXQLIRUPO\&1ODEHOHGSURWHLQXVLQJTXDOLWDWLYHGLVWDQFHUHVWUDLQWVIURP0$6VROLG

㻌

VWDWH & 105 REVHUYHG SDUDPDJQHWLFUHOD[DWLRQ HQKDQFHPHQW7DPDNL0 (JDZD $.LGR . .DPHGD 7.DPL\D0.LNXNDZD7$L]DZD7)XMLZDUD7DQG'HPXUD0  -%LRPRO105


&ORVHGF\FOH FROG KHOLXP PDJLFDQJOH VSLQQLQJ IRU VHQVLWLYLW\HQKDQFHG PXOWLGLPHQVLRQDO VROLGVWDWH 105
0DWVXNL<1DNDPXUD6)XNXL66XHPDWVX+DQG)XMLZDUD7  -0DJQ5HVRQ 



8WLOL]DWLRQ RI SDUDPDJQHWLF UHOD[DWLRQ HQKDQFHPHQWV IRU KLJKUHVROXWLRQ 105 VWUXFWXUH GHWHUPLQDWLRQ RI D
VROXEOH ORRSULFKSURWHLQ ZLWK VSDUVH 12( GLVWDQFH UHVWUDLQWV )XUXLWD . .DWDRND 6 6XJLNL 7 +DWWRUL <.RED\DVKL1,NHJDPL76KLR]DNL.)XMLZDUD7DQG.RMLPD&  -%LRPRO105



8WLOL]DWLRQRIO\VLQH&PHWK\ODWLRQ105IRUSURWHLQSURWHLQLQWHUDFWLRQVWXGLHV+DWWRUL<)XUXLWD.2KNL,

6.

6HFRQGDU\ VWUXFWXUDO DQDO\VLV RI SURWHLQV EDVHG RQ & FKHPLFDO VKLIW DVVLJQPHQWV LQ XQUHVROYHG VROLGVWDWH,NHJDPL7)XNDGD+6KLUDNDZD0)XMLZDUD7.RMLPD&  -%LRPRO105 
105VSHFWUDHQKDQFHGE\IUDJPHQWHGVWUXFWXUHGDWDEDVH,NHGD.(JDZD$)XMLZDUD7   -%LRPRO

105㻌◦◦◦ 21 ◦IPR㻌/DERUDWRU\ RI 3URWHLQ &U\VWDOORJUDSK\
㻌/DERUDWRU\RI3URWHLQ&U\VWDOORJUDSK\㻌㻌㻌
3URIHVVRU*HQML.XULVX3K'
JNXULVX#SURWHLQRVDNDXDFMS $VVRFLDWH3URIHVVRU  +LGHDNL7DQDND3K'

6SHFLDOO\DSSRLQWHG$VVRFLDWH3URIHVVRU &KULVWRSK*HUOH3K'
$VVLVWDQW3URIHVVRU  

 7HWVXNR1DNDQLZD3K'%LRORJLFDO UHDFWLRQV LQ WKH FHOO DUH FRQQHFWHG HDFK RWKHU WR IRUP D EDODQFHG QHWZRUN 3URWHLQFU\VWDOORJUDSK\ LV WKH EHVW PHWKRG WR GHWHUPLQH WKH DWRPLF VWUXFWXUH RI SURWHLQ PROHFXOHV LQ RUGHU WR

HOXFLGDWH WKH PROHFXODU PHFKDQLVP RI KLJKO\ RUJDQL]HG ELRORJLFDO V\VWHPV 7KUHHGLPHQVLRQDO SURWHLQVWUXFWXUHVHVSHFLDOO\LQSKRWRV\QWKHVLVUHVSLUDWLRQDQGF\WRVNHOHWDOPROHFXODUPRWRUVEULQJXVDGHHSHULQVLJKWLQWRWKHQHWZRUNLQJKLJKO\RUJDQL]HGELRORJLFDOSURFHVVHVLQWKHFHOO7KHPDLQDLPRIRXUUHVHDUFKJURXS LV ;UD\ VWUXFWXUH GHWHUPLQDWLRQ RI ELRORJLFDO PDFURPROHFXODU DVVHPEOLHV LQFOXGLQJ PHPEUDQH
SURWHLQ FRPSOH[HV DW WKH DWRPLF OHYHO &XUUHQW 5HVHDUFK 3URMHFWV DUH n6WUXFWXUDO VWXGLHV RI D

SKRWRV\QWKHWLF PHPEUDQHSURWHLQFRPSOH[VXFKDV&\WRFKURPH EIDQG3KRWRV\VWHP,FRPSOH[HVDQG
UHODWHGUHGR[HQ]\PHV|n&U\VWDOVWUXFWXUHDQDO\VHVRIG\QHLQPRWRUVZLWKFU\VWDOORJUDSKLFWHFKQLTXHDVZHOODV&'DQG105VSHFWURVFRSLFPHWKRGV|DQGn+LJKUHVROXWLRQDQGUDGLDWLRQGDPDJHIUHHVWUXFWXUHDQDO\VLVRIPHWDOORSURWHLQVXVLQJ;UD\IUHHHOHFWURQODVHURUQHXWURQDVDSUREH|㻌

5HVHDUFK3URJUDPV㻌

6WUXFWXUDO VWXGLHV RI SKRWRV\QWKHWLF HQHUJ\WUDQVGXFLQJ PHPEUDQH SURWHLQ FRPSOH[ DQG UHODWHG㻌&U\VWDOVWUXFWXUHDQDO\VHVRIG\QHLQPRWRUVUHGR[HQ]\PHV+LJKUHVROXWLRQDQGUDGLDWLRQGDPDJHIUHHVWUXFWXUDODQDO\VLVRIPHWDOORSURWHLQV

5HVHDUFK7RSLFV‘ '\QHLQ LV D PLFURWXEXOHEDVHG PRWRU SURWHLQ FRQVLVWLQJ RI

LGHQWLFDOKHDY\FKDLQVZLWKDVVRUWHGOLJKWOLJKWLQWHUPHGLDWHDQG

㻌

LQWHUPHGLDWH FKDLQV  0RWRU DFWLYLW\LV ORFDWHG LQ WKH KHDY\ FKDLQ㻌

WKHPROHFXODUPDVVRIZKLFKLVPRUHWKDQN'D+LJKUHVROXWLRQ㻌PROHFXODUVWUXFWXUHVLQFOXGLQJRXU;UD\VWUXFWXUH 3'%,'9.+

㻌LQGLFDWHGWKDWWKHPLFURWXEXOHELQGLQJGRPDLQRIG\QHLQKHDY\FKDLQ

LVVHSDUDWHGIURPWKH$$$FRUHRIWKHPRWRUZKLFKFRQWDLQVWKH$73

㻌

K\GURO\VLVVLWHVE\DQHORQJDWHGVWDONGRPDLQFRQVLVWLQJRIDQDQWL㻌SDUDOOHO FRLOHGFRLO㻌VWUXFWXUH )LJ  ,W ZDV K\SRWKHVL]HG WKDW WKH
㻌G\QHLQXWLOL]HGVPDOODPRXQWVRIKHOL[VOLGLQJEHWZHHQWKH$$$FRUH

㻌DQG WKH PLFURWXEXOHELQGLQJ GRPDLQ %DVHG RQ WKH FXUUHQWO\

DYDLODEOH ;UD\ VWUXFWXUHV RI VWDON FRLOHGFRLO UHJLRQV DQG UHODWHG㻌ELRSK\VLFDOFKDUDFWHUL]DWLRQVXFKDVPROHFXODUG\QDPLFVVLPXODWLRQ㻌ZHSURSRVHGWKHn2SHQ]LSSHU0RGHO|RIKHOL[VOLGLQJ+RZHYHUWKH
㻌VWUXFWXUDOEDVLVRIKRZWRVOLGHWKHWZRORQJFRLOHGFRLOKHOLFHVLQWKHRSSRVLWH GLUHFWLRQV DQG FRXSOH WKH PLFURWXEXOH ELQGLQJ LV VWLOO㻌XQNQRZQ◦ 22 ◦◦◦ Division of Protein Structural Biology)LJ2YHUDOOVWUXFWXUHRIWKHG\QHLQPRWRU
GRPDLQDW$'3ERXQGIRUP$ULEERQPRGHOVKRZLQJOLQNHU EOXH VL[$$$
PRGXOHVFRQVWLWXWLQJULQJ IURPEOXHWRUHG 
VWDONtVWUXWFRLOHGFRLOV \HOORZDQGRUDQJH DQG
&VHTXHQFH SLQN DWWKHRXWVLGHRIWKHULQJInstitute for Protein Research Osaka University㻌஁പ࣯ߑଆਫ਼෼ָ‫ڂݜ‬෨໵
஁
പ࣯ߑ
࣯ߑଆਫ਼
෼ָ‫ڂݜ‬෨໵
‫ڂ‬෨໵
㻌ଆਫ਼෼ָ
‘ /LIH RQ (DUWK GHSHQGV XOWLPDWHO\ RQ VXQOLJKW WKURXJKSKRWRV\QWKHVLV 3KRWRV\QWKHWLF HOHFWURQ WUDQVIHU FRQVLVWV RI

WKUHH PHPEUDQH SURWHLQ FRPSOH[HV 3KRWRV\WHP ,,&\WRFKURPH EIFRPSOH[DQG3KRWRV\VWHP, 36, ZKLFKDUH㻌㻌㻌

HOHFWULFDOO\FRQQHFWHGE\WKHVROXEOHHOHFWURQFDUULHUSURWHLQV㻌FKDOOHQJHWRSUHGLFWWKHSURGXFWLYHHOHFWURQWUDQVIHUFRPSOH[㻌

3ODVWRF\DQLQRU)HUUHGR[LQ )G +RZHYHULWUHPDLQVDPDMRUIRUPDWLRQ EHWZHHQ WKH PHPEUDQH SURWHLQ DQG WKH VROXEOH

HOHFWURQFDUULHUSURWHLQDQGWRXQGHUVWDQGKRZWKHUHGXFLQJHTXLYDOHQWVDUHGHOLYHUHGIURPWKHWK\ODNRLGPHPEUDQHWRWKH

㻌
㻌

㻌
㻌

VWURPDOVSDFHYLD)G2XUJURXSGHVFULEHGWKH;UD\VWUXFWXUH

RIWKHWULPHULF36,)GFRPSOH[DQGLWVFRPSOHPHQWDU\DQDO\VLV

)LJ2YHUDOOVWUXFWXUHRIWKHF\DQREDFWHULDO36,
J  2YHUDOO VWUXFWXUH RI WKH F\DQREDFWHULDO 36,)GFRPSOH[IURP7HORQJDWXV $F\WRVROLFYLHZRIWKH36,)GFRPSOH[7KUHH)GVFRORUHGLQVN\EOXH\HOORZDQGPDJHQWDDUHERXQG
WRUHVSHFWLYHSURPRWHUVRI36,WULPHU

LQ VROXWLRQ E\ 105 DQG IODVK DEVRUSWLRQ VSHFWURVFRS\ :H

GLVFRYHUHG WKDW WKH )GERXQG 36, WULPHU LV DV\PPHWULF DQGWKH SLVWRQOLNH VKLIW RI WKH WUDQVPHPEUDQH VXEXQLW 3VD)
ZKLFKLPSO\WKHPROHFXODUVLJQDOLQJDFURVVWKHPHPEUDQHWRDFFRPSOLVKHIILFLHQWHOHFWURQWUDQVIHU )LJ ‘ 7KH FKDUDFWHULVWLFSURSHUW\RISODQWW\SH)G )G LVDYHU\㻌㻌㻌

QHJDWLYHUHGR[SRWHQWLDO P9IRUPDL]HOHDI)G WKHUHIRUH

㻌LWLVSXULILHGDVDQR[LGL]HG IRUPWKURXJKQRUPDOSXULILFDWLRQ㻌SURFHGXUH$OWKRXJKLWFDQEHFKHPLFDOO\UHGXFHGDVROXWLRQRUFU\VWDOVDPSOHLVLPPHGLDWHO\R[LGL]HGE\PROHFXODUR[\JHQ㻌

0RVWRIWKHUHSRUWHG;UD\VWUXFWXUHRI)GDQG)GGHSHQGHQW㻌HQ]\PHVDUHEHOLHYHGWREHLQR[LGL]HGIRUP )LJ +RZHYHU㻌WKHVWURQJ;UD\EHDPDW6\QFKURWURQPD\SDUWLDOO\UHGXFHWKH
㻌FU\VWDOVGXULQJ;UD\EHDPLUUDGLDWLRQ,QRUGHUWRHOXFLGDWHWKH

UDGLDWLRQIUHH VWUXFWXUH RI )G DQG )GGHSHQGHQW HQ]\PHV㻌IXUWKHUGLIIUDFWLRQVWXG\RIWKHVHSURWHLQVLVQHHGHG㻌

㻌3XEOLFDWLRQV㻌)LJ6FKHPDWLFUHSUHVHQWDWLRQVRIVHPLV\QWKHWLF
>)H)H@K\GURJHQDVH WKDW FDWDO\]HV )GGHSHQGHQWK\GURJHQ SURGXFWLRQ ZLWK WKH KHOS RI PHWDOOLF
FRIDFWRUDWRPV FRORUHGVSKHUHV ;UD\ VWUXFWXUH RI DQ DV\PPHWULFDO WULPHULF IHUUHGR[LQSKRWRV\VWHP , FRPSOH[㻌 .XERWDNDZDL + 0XWRK 5

6KLQPXUD.6ËWLI31RZDF]\N005ØJQHU0,NHJDPL77DQDND+DQG.XULVX*  1DWXUH3ODQWV

&DOUHGR[LQUHSUHVHQWVDQRYHOW\SHRIFDOFLXPGHSHQGHQWVHQVRUUHVSRQGHUFRQQHFWHGWRUHGR[UHJXODWLRQLQWKH
FKORURSODVW+RFKPDO$.=LQ]LXV.&KDURHQZDWWDQDVDWLHQ5*ÇEHOHLQ30XWRK57DQDND+6FKXO]H

6/LX*6FKRO]01RUGKXHV$2IIHQERUQ-13HWURXWVRV')LQD]]L*)XIH]DQ&+XDQJ..XULVX





*DQG+LSSOHU0  1DWXUH&RPPXQ$VWUXFWXUDOYLHZRIV\QWKHWLFFRIDFWRULQWHJUDWLRQLQWR>)H)H@K\GURJHQDVHV(VVHOERUQ-0XUDNL1.OHLQ.
(QJHOEUHFKW90HW]OHU1ROWH1$SIHO83+RIPDQQ(.XULVX*DQG+DSSH7   &KHP6FL



7KH«FU\VWDOVWUXFWXUHRIWKHG\QHLQPRWRUGRPDLQ.RQ72\DPD76KLPR.RQ5,PDPXOD.6KLPD

76XWRK.DQG.XULVX*  1DWXUH

;UD\FU\VWDOVWUXFWXUHRIWKHOLJKWLQGHSHQGHQWSURWRFKORURSK\OOLGHUHGXFWDVH0XUDNL11RPDWD-(EDWD(0L]RJXFKL76KLED77DPLDNL+.XULVX*DQG)XMLWD<  1DWXUH

6WUXFWXUHRIWKH&\WRFKURPH EIFRPSOH[RI2[\JHQLF3KRWRV\QWKHVLV7XQLQJWKHFDYLW\.XULVX*=KDQJ+

6WUXFWXUHRIWKHHOHFWURQWUDQVIHUFRPSOH[EHWZHHQIHUUHGR[LQDQGIHUUHGR[LQ1$'3  UHGXFWDVH.XULVX*

6PLWK-/DQG&UDPHU:$  6FLHQFH .XVXQRNL0.DWRK(<DPD]DNL77HVKLPD.2QGD<.LPDWD$ULJD<DQG+DVH7  1DWXUH6WUXFW%LRO

◦◦◦ 23 ◦IPR㻌/DERUDWRU\ RI 0HPEUDQH 3URWHLQ &KHPLVWU\
㻌/DERUDWRU\RI0HPEUDQH3URWHLQ&KHPLVWU\㻌㻌㻌,QGHSHQGHQW$VVRFLDWH3URIHVVRU-RML0LPD3K'
-RML0LPD#SURWHLQRVDNDXDFMS 

7KH /DERUDWRU\ RI 0HPEUDQH 3URWHLQ &KHPLVWU\ LV ZRUNLQJ RQ LQ YLWUR UHFRQVWLWXWHG SURWHROLSRVRPDOVWXGLHVWRXQGHUVWDQGWKHPROHFXODUPDFKLQHULHVRIPHPEUDQHWHWKHULQJGRFNLQJDQGIXVLRQSURFHVVHVLQ WKH HQGRPHPEUDQH RUJDQHOOH V\VWHPV RI HXNDU\RWLF FHOOV IURP \HDVW WR KXPDQV  ,QWUDFHOOXODUPHPEUDQH WHWKHULQJ GRFNLQJ DQG IXVLRQ HYHQWV DUH IXQGDPHQWDO DQG FRQVHUYHG ELRORJLFDO UHDFWLRQVZKLFKDUHYLWDOIRUYHVLFOHWUDIILFNLQJEHWZHHQVXEFHOOXODURUJDQHOOHPHPEUDQHFRPSDUWPHQWVDQGSODVPDPHPEUDQHV RUJDQHOOH PRUSKRORJ\ KRUPRQH VHFUHWLRQ DQG DOVR V\QDSWLF QHXURWUDQVPLVVLRQ (DUOLHUVHPLQDOVWXGLHVKDYHUHYHDOHGWKDWPHPEUDQHWHWKHULQJGRFNLQJDQGIXVLRQDUHWHPSRUDOO\DQGVSDWLDOO\UHJXODWHGLQFHOOVE\WKHGLYHUVHVHWVRINH\SURWHLQFRPSRQHQWVZKLFKLQFOXGH61$5(IDPLO\SURWHLQV

61$5(LQWHUDFWLQJFKDSHURQHSURWHLQVVXFKDV6HF0XQFIDPLO\SURWHLQV5DEIDPLO\VPDOO*73DVHV

5DELQWHUDFWLQJ HIIHFWRU SURWHLQV DQG WHWKHULQJ PXOWLVXEXQLW FRPSOH[HV +RZHYHU LW KDV VWLOO UHPDLQHG

HQLJPDWLFKRZWKRVHHVVHQWLDOSURWHLQIDFWRUVFRRSHUDWHWRVSHFLILFDOO\DQGHIILFLHQWO\PHGLDWHPHPEUDQH

WHWKHULQJ GRFNLQJ DQG IXVLRQ HYHQWV ,Q RXU JURXS ZH KDYH EHHQ DQG FRQWLQXH WR H[SORUH WKHVH YLWDOWHWKHULQJGRFNLQJIXVLRQ PDFKLQHULHV E\ LQ YLWUR UHFRQVWLWXWLRQ ZLWK SXULILHG UHFRPELQDQW SURWHLQV

61$5(V 5DEV DQG WKHLU HIIHFWRUV  DQG V\QWKHWLF OLSLG ELOD\HUV ZLWK GHILQHG OLSLG FRPSRVLWLRQV 2XUUHFRQVWLWXWLRQ VWXGLHV ZLOO XQFRYHU WKH PHFKDQLVWLF GHWDLOV RI KRZ HXNDU\RWLF FHOOV FRQIHU WKHVSDWLRWHPSRUDOVSHFLILFLW\RIWKHHQGRPHPEUDQHWUDIILFNLQJV\VWHPV

5HVHDUFK3URJUDPV

8QGHUVWDQGLQJWKHPROHFXODUPDFKLQHU\WRFDWDO\]HPHPEUDQHWHWKHULQJGRFNLQJDQGIXVLRQHYHQWV

LQHXNDU\RWLFHQGRPHPEUDQHV\VWHPV'HYHORSLQJQRYHOUHFRQVWLWXWLRQV\VWHPVDQGDVVD\VWRVWXG\WKHPHFKDQLVPVE\ZKLFK61$5(V5DE*73DVHVDQGWKHLUHIIHFWRUVFRQWUROWKHGLUHFWLRQDOLW\RIPHPEUDQHWUDIILFNLQJ

3XEOLFDWLRQV

5HFRQVWLWXWLRQRIPHPEUDQHWHWKHULQJPHGLDWHGE\5DEIDPLO\VPDOO*73DVHV0LPD-   %LRSK\V5HY

+XPDQ 5DE VPDOO *73DVH DQG FODVV 9 P\RVLQPHGLDWHG PHPEUDQH WHWKHULQJ LQ D FKHPLFDOO\ GHILQHG
UHFRQVWLWXWLRQV\VWHP,QRVKLWD00LPD-  -%LRO&KHP



3K\VLRORJLFDO OLSLG FRPSRVLWLRQ LV YLWDO IRU KRPRW\SLF (5 PHPEUDQH IXVLRQ PHGLDWHG E\ WKH G\QDPLQUHODWHG

0HPEUDQHDQFKRUHG KXPDQ 5DE *73DVHV GLUHFWO\ PHGLDWH PHPEUDQH WHWKHULQJ LQ YLWUR 7DPXUD 1 0LPD -*73DVH6H\S6XJLXUD60LPD-  6FL5HS
 %LRO2SHQ

0XOWLSOH DQG GLVWLQFW VWUDWHJLHV RI \HDVW 61$5(V WR FRQIHU WKH VSHFLILFLW\ RI PHPEUDQH IXVLRQ )XUXNDZD 1'LVWLQFWFRQWULEXWLRQVRIYDFXRODU4DEFDQG561$5(SURWHLQVWRPHPEUDQHIXVLRQVSHFLILFLW\,]DZD52QRXH

0LPD-  6FL5HS
7)XUXNDZD10LPD-  -%LRO&KHP

5HFRQVWLWXWHGPHPEUDQHIXVLRQUHTXLUHVUHJXODWRU\OLSLGV61$5(VDQGV\QHUJLVWLF61$5(FKDSHURQHV0LPD-
+LFNH\&0;X+-XQ<:LFNQHU:  (0%2-

◦ 24 ◦◦◦ Division of Protein Structural Biology

Division of Integrated
Protein Functions

Laboratory of Genome and Chromosome Functions
Laboratory for Molecular and Developmental Biology
Laboratory for Advanced Brain Functions
Laboratory for Nuclear Dynamics
Laboratory of Homeostatic Integration

IPR
㻌

\
㻌/DERUDWRU\RI*HQRPHDQG&KURPRVRPH)XQFWLRQV㻌㻌㻌
3URIHVVRU$NLUD6KLQRKDUD'6FLDVKLQR#SURWHLQRVDNDXDFMS $VVRFLDWH3URIHVVRU   $VVLVWDQW3URIHVVRU

 $VDNR)XUXNRKUL3K'.HQLFKLUR0DWVX]DNL3K'
0LND+LJDVKLGH3K':H DUH VWXG\LQJ PHFKDQLVPV WR VWDELOL]H JHQRPH DQG PROHFXODU GHIHFWV LQGXFHG E\JHQRPLFLQVWDELOLW\E\YLVXDOL]LQJPROHFXODUSDWKZD\VDWDPROHFXODUOHYHODVZHOODVGHFLSKHULQJWKHHYHQWVLQFHOOV
RULQRUJDQLVPV$PRQJSDWKZD\VWRVWDELOL]HWKHJHQRPHZHDUHIRFXVLQJRQQRWRQO\'1$ELRFKHPLVWU\RI'1$

H[FKDQJH KRPRORJRXV UHFRPELQDWLRQ DQG QRQKRPRORJRXV HQG MRLQLQJ EXW DOVR FRQWURO PHFKDQLVPV RI WKHLUUHFRPELQDWLRQHYHQWVLQPLWRWLFDQGPHLRWLFFHOOF\FOHVE\FKURPRVRPHPRUSKRJHQHVLVDQGPRYHPHQWV:HHPSOR\HG
LQWHUGLVFLSOLQDU\ DSSURDFKHV FRPELQHG ZLWK PROHFXODU ELRORJ\ ELRFKHPLVWU\ VWUXFWXUDO ELRORJ\ DQG JHQRPHZLGHDSSURDFKHV5HFHQWO\ZHKDYHEHHQH[WHQGLQJRXUDQDO\VLVE\WDUJHWLQJUHFRPELQDWLRQUHDFWLRQVLQKXPDQFHOOVRU

PLFH'\VIXQFWLRQRIUHFRPELQDWLRQLQPLWRWLFFHOOVLQGXFHVJHQRPHLQVWDELOLW\ZKLFKOHDGVWRWXPRULJHQHVLVZKLOH

GHIHFWLYH UHFRPELQDWLRQ GXULQJ JDPHWH IRUPDWLRQ LQGXFHV PLVFDUULDJH DQG DQHXSORLG\ GLVHDVH VXFK DV 'RZQV\QGURPH:HDUHDOVRWU\LQJWRXQGHUVWDQGPROHFXODUGHIHFWVLQWKHVHGLVHDVHV

5HVHDUFK3URJUDPV

6WXG\RIIXQFWLRQVDQGVWUXFWXUHRISURWHLQDQGSURWHLQFRPSOH[HVLQYROYHGLQKRPRORJRXVUHFRPELQDWLRQ&RQWUROPHFKDQLVPVRIUHFRPELQDWLRQE\HSLJHQHWLFVDQGWHORPHUHIXQFWLRQV

6WXG\RQUHFRPELQDWLRQDQG'1$UHSDLULQPLFHDQGKXPDQFHOOV5HVHDUFKRQLQWHUSOD\EHWZHHQUHFRPELQDWLRQDQGFKURPRVRPHVWUXFWXUHPRWLRQDQGQXFOHDUHQYHORSHUHPRGHOLQJ

0HFKDQLVPRISDWKZD\FKRLFHLQGRXEOHVWUDQGEUHDN '$% UHSDLUGXULQJFHOOF\FOHRIKXPDQFHOO

5HVHDUFK7RSLFV‘,QUHFRPELQDWLRQDVWHSFDOOHGKRPRORJVHDUFKEHWZHHQ'1$VLVDNH\VWHSLQYROYLQJG\QDPLFDVVHPEO\DQGGLVDVVHPEO\RISURWHLQVRQVLQJOHVWUDQGHG
VV '1$$5HF$KRPRORJ5DGIRUPVDULJKWKDQGHGKHOLFDOILODPHQWRQ
VV'1$ DVVHPEO\ DQG GLVDVVHPEO\ RI ZKLFK LV WLJKWO\ UHJXODWHG E\ ERWKSRVLWLYH DQG QHJDWLYHUHJXODWRUV 5DGSDUDORJV DV ZHOODV %5&$ ZKRVHPXWDWLRQ FDXVHV IDPLOLDO EUHDVWFDQFHU WR SURPRWH WKH IRUPDWLRQ RI 5DGILODPHQW +RZHYHU KRZ WKHVH SURWHLQV ZRUN LQ 5DG DVVHPEO\ UHPDLQVXQNQRZQ :H KDYH EHHQ VWXG\LQJ 5DGPHGDWHG UHDFWLRQ VLQFH ZH
LGHQWLILHG 5DG DV D 5HF$ KRPRORJ 5HFHQWO\ ZH LGHQWLILHG D QHZ SURWHLQFRPSOH[ FDOOHG 3&66  ZKLFK IDFLOLWDWHV 5DGDVVHPEO\DQG GHWHUPLQHV D
FU\VWDOVWUXFWXUHRIFRUHFRPSOH[RI3V\&VP7KHVHVWXGLHVVKHGOLJKWRQ
KRZWKLVFRPSOH[SURPRWHV5DGILODPHQWE\ELQGLQJWRWKHHQGRIILODPHQW
IRUUHFUXLWPHQWDVZHOODVVWDELOL]DWLRQ 5HI ‘'XULQJ PHLRVLV VSHFLDOL]HG FKURPRVRPH PRUSKRJHQHVLV VXFK DV WKH
IRUPDWLRQRIWKH6\QDQSWRQHPDOFRPSOH[ 6& DQGG\QDPLFVZLWKWHORPHUH
FOXVWHULQJDUHEHOLHYHGWRFRQWURO'1$ELRFKHPLVWU\RQPHLRWLFFKURPRVRPHVVXFKDVUHFRPELQDWLRQ,WUHPDLQVXQVROYHGKRZWKH6&LVIRUPHGWHORPHUH
FOXVWHULQJLVIDFLOLWDWHGLQWHUPVRIPROHFXODUPDFKLQHU\2XUJURXSKDVEHHQ
LGHQWLI\LQJ SURWHLQV LQYROYHG LQ WKHVH SURFHVVHV 5HFHQWO\ ZH KDYH EHHQIRFXVLQJRQWKHFRKHVLQFRPSOH[HVLQYROYHGLQVLVWHUFKURPDWLGFRKHVLRQLQ㻌PHLRWLFHYHQWV DQG IRXQG D UHJXODWRU RI FRKHVLQ SOD\V D FULWLFDO UROH LQ WKH
FRPSDFWLRQ RI FKURPRVRPH D[HV DQG FRQWUROV WHORPHUH G\QDPLFV E\
PHGLDWLQJUHPRGHOLQJRIQXFOHDUHQYHORSHV 5HI

◦ 26 ◦◦◦ Division of Integrated Protein Functions$PRGHORI5DGDVVHPEO\,PPXQRVWDLQLQJ LPDJH RI 5HG JUHHQ 
DQG5HFFRKHVLQ UHG Institute for Protein Research Osaka University
㻌‘0HLRWLFUHFRPELQDWLRQSOD\VDQHVVHQWLDOUROHLQFKURPRVRPHVHJUHJDWLRQ㻌㻌DWPHLRVLV,'LIIHUHQWIURPPLWRWLFUHFRPELQDWLRQPHLRWLFUHFRPELQDWLRQLVKLJKO\
GLIIHUHQWLDWHG DQG UHJXODWHG VR WKDW UHFRPELQDWLRQ RFFXUV㻌SUHIHUHQWLDOO\ EHWZHHQ KRPRORJRXV FKURPRVRPHV RYHU D VLVWHU FKURPDWLG㻌FURVVRYHULVPRUHIUHTXHQWWKDQQRQFURVVRYHUVWKHQXPEHUDQGGLVWULEXWLRQ㻌 FURVVRYHUV DORQJ FKURPRVRPHV DUH VWULFWO\ FRQWUROOHG (DFK XQLTXH
RI
SURSHUW\RIPHLRWLFUHFRPELQDWLRQLVFDWDO\]HGE\GLIIHUHQWSURWHLQFRPSOH[HV㻌
2XUJURXSKDVLVRODWHGGLIIHUHQWSURWHLQVLQYROYHGLQWKHVHUHDFWLRQV5HFHQW㻌VWXGLHV UHYHDOHG WKDW '1$ GDPDJH UHVSRQVH ''5 SURWHLQV VXFK DV '1$
GDPDJHFODPSSURPRWHVFURVVRYHUIRUPDWLRQE\KHOSLQJWKHDVVHPEO\RIVR㻌
FDOOHGSURFURVVRYHUFRPSOH[VXJJHVWLQJLQWHUSOD\EHWZHHQ''5DQGPHLRWLF㻌UHFRPELQDWLRQ 5HI ,PPXQRVWDLQLQJLPDJHRI6&=LS UHG 5HF JUHHQ 㻌㻌 ,W LV NQRZQ WKDWRQH RI WKH UHDVRQVZK\ FHOOV EHFRPH FDQFHURXVLV WKDW‘JHQRPLF'1$WKHVRXUFHRIRXUJHQHWLFLQIRUPDWLRQLVSK\VLFDOO\GHVWUR\HG㻌
7KXV '1$ GDPDJH RIWHQ FDXVHV JHQRPLF LQVWDELOLW\ VXFK DV PXWDWLRQ㻌
WUDQVORFDWLRQDPSOLILFDWLRQRU GHOHWLRQ ,RQL]LQJ UDGLDWLRQ VRPH FKHPLFDOVVXQOLJKWDQG
R[\JHQ
DUH
HVVHQWLDO
IRU RXU OLIH WKH\ DOVR LQGXFH JHQRPLF
㻌
LQVWDELOLW\7KHUHIRUHZHKDYHPXOWLSOHPHFKDQLVPVWRUHSDLU'1$GDPDJH
㻌
LW KDV QRW EHHQ FODULILHG ZK\ D QRUPDO FHOO WXUQV LQWR D FDQFHU FHOO RU ZK\㻌LRQL]LQJUDGLDWLRQH[SRVXUHFDXVHVFDQFHU2XUUHVHDUFKJRDOLVWRXQGHUVWDQG
WKHLQLWLDOVWDJHRIFHOOWXPRULJHQHVLVFDXVHGE\'1$GDPDJHE\XVLQJKXPDQ㻌
FHOOOLQHVGHULYHGIURPSDWLHQWVZLWKDQLQKHULWHGGLVRUGHUZLWKDKLJKULVNRI㻌
FDQFHU RU KXPDQ FHOO OLQHV ZLWK PXWDWLRQV LQWURGXFHG E\ D JHQH HGLWLQJWHFKQLTXHDQGWKHLU\HDVWPRGHOV:HUHSRUWHG
WKDWLQWKH\HDVWPRGHORI㻌KXPDQGLVRUGHUUHSDLUSURWHLQVZKLFKVKRXOGEHUHPRYHGLQVWHDGUHPDLQDQG
㻌D ORVV RI JHQHWLF LQIRUPDWLRQ FDQ EH SURYRNHG ZKLFK ZKHQ UHSDLUHG㻌
LQFRUUHFWO\ZLOOOHDGWRWKHWXPRUIRUPDWLRQDQGLPPXQRGHILFLHQF\
 5HI - Etoposide+ Etoposidefragmented
fragmented

dicentric-Etp
dicentric
fragmented

7HUDVDZD0HWDO&DQFHU6FL
'1$ GDPDJH LQGXFHG JHQRPLF LQVWDELOLW\0LWRWLF FKURPRVRPH IURP KXPDQ FHOOZLWKRXW '1$ GDPDJH OHIW  RU DIWHU '1$
GDPDJHLQGXFWLRQ ULJKW  3XEOLFDWLRQV㻌

7KH 05; &RPSOH[ (QVXUHV 1+(- )LGHOLW\ WKURXJK 0XOWLSOH 3DWKZD\V ,QFOXGLQJ;UV)+$'HSHQGHQW 7HO㻌$FWLYDWLRQ ,ZDVDNL ' +D\DVKLKDUD . 6KLPD + 7HUDVDZD0 +LJDVKLGH 0 6XVDQ * DQG 6KLQRKDUD 0 3/R6*HQHWLFVH
5DG:SO :DSO DFRKHVLQUHJXODWRUFRQWUROVFKURPRVRPHFRPSDFWLRQGXULQJPHLRVLV  1XFOHLF$FLGV
5HVHDUFK

7KH3DI&RPSOH[6KDSHVWKH/DQGVFDSHRI'RXEOHVWUDQG%UHDNVDORQJ0HLRWLF&KURPRVRPHVLQ6DFFKDURP\FHV
FHUHYLVLDH 6DQWRVK * . 3DWHO .- &ROOHWWL 00 6DVDQXPD + 6KLQRKDUD 0 +RFKZDJHQ $ DQG $
6KLQRKDUD*HQHWLFV

'1$ GDPDJH UHVSRQVH FODPS FRQWULEXWHVWR FKURPRVRPDODVVHPEO\ RI =006,& SURFURVVRYHUIDFWRUVGXULQJPHLRVLV6KLQRKDUD0+D\DVKLKDUD.*UXEE-7%LVKRS'.DQG6KLQRKDUD$  -&HOO6FL
&DQRQLFDO QRQKRPRORJRXV HQG MRLQLQJ LQ PLWRVLV LQGXFHV JHQRPH LQVWDELOLW\ DQG LV VXSSUHVVHG E\ 0SKDVHVSHFLILF SKRVSKRU\ODWLRQ RI ;5&& YLD &'.V 7HUDVDZD 0 6KLQRKDUD $ DQG 6KLQRKDUD 0   3/R6
*HQHWLFVH
$ QHZ SURWHLQ FRPSOH[ SURPRWLQJ WKH DVVHPEO\ RI 5DG ILODPHQWV 6DVDQXPD + 7DZDUDPRWR 06 /DR -
+RVDND+6DQGD(6X]XNL0<DPDVKLWD(6KLQRKDUD0+XQWHU11DNDJDZD$DQG6KLQRKDUD$  
1DWXUH&RPPXQ

7KH0SV681GRPDLQLVLPSRUWDQWIRUFKURPRVRPHPRWLRQDQGMX[WDSRVLWLRQRIKRPRORJRXVFKURPRVRPHVGXULQJPHLRVLV5DR+%'36KLQRKDUD0DQG$6KLQRKDUD*HQHVWR&HOOV
6DFFKDURP\FHVFHUHYLVLDH6UVKHOLFDVHGLVDVVHPEOHV5DGIURPPHLRWLFFKURPRVRPHV6DVDQXPD+)XULKDWD
<6KLQRKDUD0DQG6KLQRKDUD$  *HQHWLFV

&URVVRYHU DVVXUDQFH DQG FURVVRYHU LQWHUIHUHQFH DUH GLVWLQFWO\ UHJXODWHG E\ WKH =00 SURWHLQV GXULQJ \HDVWPHLRVLV6KLQRKDUD02K6+XQWHU1DQG6KLQRKDUD$  1DWXUH*HQHW  $SURWHLQFRPSOH[FRQWDLQLQJ0HLDQG6DHSURPRWHVWKHDVVHPEO\RIWKHPHLRVLVVSHFLILF5HF$KRPRORJ'PF+D\DVH$7DNDJL00L\D]DNL72VKLXPL+6KLQRKDUD0DQG6KLQRKDUD$  &HOO㻌◦◦◦ 27 ◦

IPR
㻌

\
S
J\
㻌/DERUDWRU\IRU0ROHFXODUDQG'HYHORSPHQWDO%LRORJ\㻌㻌
3URIHVVRU

7DNDKLVD)XUXNDZD0'3K'
WDNDKLVDIXUXNDZD#SURWHLQRVDNDXDFMS $VVRFLDWH3URIHVVRU<RVKLKLUR2PRUL0'3K'$VVLVWDQW3URIHVVRU

7DUR&KD\D0'3K'   $VVLVWDQW3URIHVVRU<XNR6XJLWD0'3K'

2XUODERUDWRU\VWXGLHVPROHFXODUPHFKDQLVPVXQGHUO\LQJWKHGHY HORSPHQWDQGIXQFWLRQRIWKHYHUWHEUDWH

FHQWUDO QHUYRXV V\VWHP &16  XVLQJ YDULRXV UHVHDUFK PHWKRGV RI PROHFXODU ELRORJ\ PRXVH JHQHWLFVELRFKHPLVWU\FHOOELRORJ\DQGQHXUDOSK\VLRORJ\2XUEUDLQFRQVLVWVRIPRUHWKDQELOOLRQQHXURQV7R

IXQFWLRQDVDEUDLQQXPHURXVQXPEHUVRIQHXURQVPXVWEHJHQHUDWHGDWWKHULJKWSODFHDQGWKH\PXVWEH

LQWHUFRQQHFWHG HDFK RWKHU :H XVH WKH UHWLQD DV D PRGHO V\VWHP WR XQGHUVWDQG KRZ '1$ HQFRGHVSURJUDPV WR JHQHUDWH YDULRXV QHXURQV DQG JOLDO FHOOV IRUP SUHFLVH QHXURQDO FLUFXLWV DQG HQDEOHFRPSOLFDWHGQHXURQDOIXQFWLRQ:HDOVRIRFXVRQKRZDEQRUPDOLW\RIELRORJLFDOSURFHVVHVLQGHYHORSPHQW

DQGPDWXUDWLRQOHDGVWRKXPDQGLVHDVHV:HDUHHDJHUWRFRQWULEXWHWRWKHGHYHORSPHQWRIGLDJQRVLVDQG
FXUHRIKXPDQGLVHDVHV7RJHWKHURXUODEDLPVWR HOXFLGDWHWKHPHFKDQLVPVDQGSULQFLSOHVXQGHUO\LQJ

&16GHYHORSPHQWIURP'1$SURJUDPVWRSK\VLRORJLFDOIXQFWLRQDQGKXPDQGLVHDVH

5HVHDUFK3URJUDPV$QDO\VLVRIPROHFXODUPHFKDQLVPVXQGHUO\LQJQHXURQDOGLIIHUHQWLDWLRQ$QDO\VLVRIIXQFWLRQDOPHFKDQLVPVRIWKHUHWLQDO212))QHXURQDOSDWKZD\V








0ROHFXODUDQDO\VLVRIQHXUDOFLUFXLWIRUPDWLRQLQWKHUHWLQD(OXFLGDWLRQRIIXQFWLRQDOUROHVRIPLFUR51$V PL51$V LQ&16GHYHORSPHQW)XQFWLRQDODQDO\VLVRIFLOLXPDQDQWHQQDRIDFHOOLQWKH&16$QDO\VLVRIYLVXDOIXQFWLRQXVLQJPRXVHPRGHOV

5HVHDUFK7RSLFV‘([TXLVLWHO\SUHFLVHV\QDSVHIRUPDWLRQLVFUXFLDOIRUFRUUHFWIXQFWLRQLQJRIWKHPDPPDOLDQFHQWUDOQHUYRXVV\VWHP

5HWLQDO SKRWRUHFHSWRUV WUDQVIHU LQIRUPDWLRQ WR ELSRODU DQG KRUL]RQWDO FHOOV DW D VSHFLDOL]HG V\QDSVH WKH ULEERQV\QDSVH:HLGHQWLILHGSLNDFKXULQDQRYHOH[WUDFHOOXODUPDWUL[OLNHUHWLQDOSURWHLQDQGREVHUYHGLWVORFDOL]DWLRQWRWKH

V\QDSWLFFOHIWLQWKHSKRWRUHFHSWRUULEERQV\QDSVH 3LNDFKXULQQXOOPXWDQWPLFHGLVSOD\LPSURSHUDSSRVLWLRQRIWKH

ELSRODUWHUPLQXVWRWKHULEERQV\QDSVHUHVXOWLQJLQDOWHUDWLRQVLQV\QDSWLFVLJQDOWUDQVPLVVLRQDQGYLVXDOIXQFWLRQDV

GHWHFWHGE\HOHFWURUHWLQRJUDPDQGRSWRNLQHWLFUHVSRQVHV:HREVHUYHGFRORFDOL]DWLRQRISLNDFKXULQZLWKG\VWURSKLQJO\FRSURWHLQ FRPSOH[ '*&  PROHFXOHV DQG GLUHFW ELQGLQJ RI SLNDFKXULQ ZLWK DOSKDG\VWURJO\FDQ DQ H[WUDFHOOXODU㻌FRPSRQHQWRI'*&7RJHWKHURXUUHVXOWVLGHQWLILHGDQRYHOG\VWURJO\FDQOLJDQGSURWHLQSLNDFKXULQDQGGHPRQVWUDWH
LWVHVVHQWLDOUROHLQWKHSUHFLVHLQWHUDFWLRQVEHWZHHQWKHSKRWRUHFHSWRUULEERQV\QDSVHDQGWKHELSRODUWHUPLQXV7KLV㻌
PD\ DOVR DGYDQFH RXU XQGHUVWDQGLQJ RI WKH PROHFXODU PHFKDQLVPV XQGHUO\LQJ WKH UHWLQDO HOHFWURSK\VLRORJLFDODEQRUPDOLWLHVREVHUYHGLQPXVFXODUG\VWURSK\SDWLHQWV 5HI )LJ (OHFWURQ WRPRJUDSK\ RI URG SKRWRUHFHSWRU V\QDSVHWHUPLQDOVXVLQJXOWUDKLJKYROWDJHHOHFWURQPLFURVFRS\,QWKH:7PRXVHUHWLQDSKRWRUHFHSWRUD[RQDO WHUPLQXVIRUPVLQYDJLQDWLRQWRDSSRVHGHQGULWLFWHUPLQDOVRIKRUL]RQWDOFHOOV
DQG ELSRODU FHOOV WR IRUP WKH ULEERQ V\QDSVH ,Q FRQWUDVW3LNDFKXULQ .2 PLFH VKRZHG LPSURSHU DSSRVLWLRQ RI WKHELSRODU FHOO GHQGULWLF WLSV WR WKH SKRWRUHFHSWRU ULEERQV\QDSVHV◦ 28 ◦◦◦ Division of Integrated Protein Functions

Institute for Protein Research Osaka University‘ 0LFUR51$LVDERXWESVPDOOQRQFRGLQJ51$ZKLFKLVLQYROYHGLQ

PDQ\ ELRORJLFDO SURFHVVHV DQG KXPDQ GLVHDVHV WKURXJK WUDQVODWLRQDO

UHSUHVVLRQ 0LFUR51$D PL5D  LV WKH PRVW DEXQGDQW PL51$

H[SUHVVHGLQWKHYHUWHEUDWHFHQWUDOQHUYRXVV\VWHP &16 :HH[SORUHG

WKHLQYLYRIXQFWLRQRIPL5DE\WDUJHWHGGLVUXSWLRQRI 5QFU UHWLQDO㻌QRQFRGLQJ51$ ZKLFKLVWKHGRPLQDQWVRXUFHRIPL5D5QFUtt
PXWDQWPLFHH[KLELWDEQRUPDOLWLHVLQWKH&16LQFOXGLQJVPDOOEUDLQVL]HD[RQDOPLVVSURXWLQJRI'*JUDQXOHFHOOVDQGUHWLQDOFRQHFHOOGHDWK:H

IRXQGWKDW/K[LVDQLQYLYRWDUJHWP51$RIPL5D:HDOVRREVHUYHG

WKDW /+; GRZQUHJXODWLRQ E\ PL5D LV UHTXLUHG IRU SUHYHQWLRQ RIDSRSWRVLV LQ WKH GHYHORSLQJ UHWLQD DQG SURSHU D[RQDO GHYHORSPHQW RI

KLSSRFDPSDO QHXURQV 2XU UHVXOWV VKRZHG WKDW PL5D SOD\V DQ

HVVHQWLDOUROHLQPDWXUDWLRQDQGVXUYLYDORI'*QHXURQVDQGUHWLQDOFRQHV
E\UHSUHVVLQJ/K[WUDQVODWLRQ 5HI 㻌)LJ$EHUUDQWVSURXWLQJRIPRVV\ILEHUVLQWKHPL5D.2PRXVH
7KHPRVV\JLYHUWHUPLQDOVZHUHYLVXDOL]HGE\7LPPVWDLQLQJZLWK1LVVO
FRXQWHUVWDLQLQJDWSRVWQDWDOGD\6FDOHEDUVUHSUHVHQWXP

‘&LOLDDUHPLFURWXEXOHEDVHGRUJDQHOOHVWKDWH[WHQGIURPWKHVXUIDFHRIYDULRXVW\SHVRIFHOOVLQ YHUWHEUDWHV ,Q KXPDQV G\VIXQFWLRQ RI FLOLD FDXVHV D EURDG UDQJH RI RYHUODSSLQJ FOLQLFDOSKHQRW\SHV WHUPHG nFLOLRSDWKLHV| ZKLFK LQFOXGH UHWLQDO GHJHQHUDWLRQ SRO\F\VWLF NLGQH\

GLVHDVH SRO\GDFW\O\ DQG REHVLW\ :H IRXQG WKDW D FLOLDU\ NLQDVH 0DN LV HVVHQWLDO IRU FLOLDU\OHQJWKUHJXODWLRQDQGVXUYLYDORISKRWRUHFHSWRUFHOOV 5HI :HFODULILHGWKDWD0DNSDUDORJ

,&.UHJXODWHVWXUQDURXQGPDFKLQHU\RILQWUDIODJHOODUWUDQVSRUW ,)7 DWFLOLDU\WLSVXVLQJNQRFNRXWPLFHDQG]HEUDILVKPRUSKDQWV 5HI )LJ$K\SRWKHWLFDOPRGHOIRU,&.IXQFWLRQLQWKHUHJXODWLRQRI
,)37WXUQDURXQGDWFLOLDU\WLSV ,&. UHJXODWHV WKH GLVDVVHPEO\ EHWZHHQ ,)7$ DQG ,)7%
VXEFRPSOH[HVDWFLOLDU\WLSVWKURXJKSKRVSKRU\ODWLRQRI,)7

3XEOLFDWLRQV






/ULWD5HWLQDO7UDQVPHPEUDQH 3URWHLQ5HJXODWHV6HOHFWLYH6\QDSVH)RUPDWLRQLQ&RQH3KRWRUHFHSWRU&HOOV

DQG9LVXDO$FXLW\8HQR$HWDO  &HOO5HSt

㻌

6DPGLVDFHOOW\SHVSHFLILF35&FRPSRQHQWHVVHQWLDOIRUHVWDEOLVKLQJUHWLQDOURGSKRWRUHFHSWRULGHQWLW\2PRUL< HWDO  3URF1DWO$FDG6FL86$((

3URWHLQ*0HGLDWHG0HPEUDQH7UDIILFNLQJ,V(VVHQWLDOIRU&RUUHFW5RG6\QDSWLF/RFDWLRQLQWKH5HWLQDDQG
IRU1RUPDO9LVXDO)XQFWLRQ6DQXNL5HWDO  &HOO5HSt,&. LV HVVHQWLDO IRU FHOO W\SHVSHFLILF FLOLRJHQHVLV DQG WKH UHJXODWLRQ RI FLOLDU\ WUDQVSRUW &KD\D 7 2PRUL <.XZDKDUD5)XUXNDZD7  (0%2-

3UHV\QDSWLF '\VWURJO\FDQ3LNDFKXULQ &RPSOH[ 5HJXODWHV WKH 3URSHU 6\QDSWLF &RQQHFWLRQ EHWZHHQ 5HWLQDO

3KRWRUHFHSWRUDQG%LSRODU&HOOV2PRULHWDO  -1HXURVFL 



PL5DLVUHTXLUHGIRUKLSSRFDPSDOD[RJHQHVLVDQGUHWLQDOFRQHVXUYLYDOWKURXJK/K[VXSSUHVVLRQ6DQXNL5



1HJDWLYH UHJXODWLRQRI FLOLDU\ OHQJWK E\ FLOLDU\ PDOH JHUP FHOODVVRFLDWHG NLQDVH 0DN  LV UHTXLUHG IRU UHWLQDO

HWDO  1DWXUH1HXURVFLSKRWRUHFHSWRUVXUYLYDO2PRUL<HWDO  31$6



%OLPS VXSSUHVVHV &K[ H[SUHVVLRQ LQ GLIIHUHQWLDWLQJ UHWLQDO SKRWRUHFHSWRU SUHFXUVRUV WR HQVXUH SURSHU

3LNDFKXULQDG\VWURJO\FDQOLJDQGLVHVVHQWLDOIRUSKRWRUHFHSWRUULEERQV\QDSVHIRUPDWLRQ6DWR6HWDO SKRWRUHFHSWRUGHYHORSPHQW.DWRK.HWDO  -1HXURVFL1DWXUH1HXURVFL

◦◦◦ 29 ◦IPR㻌/DERUDWRU\ IRU $GYDQFHG %UDLQ )XQFWLRQV
㻌/DERUDWRU\IRU$GYDQFHG%UDLQ)XQFWLRQV㻌㻌㻌
3URIHVVRU
7DNDWRVKL+LNLGD0'3K'ʤKLNLGDˑSURWHLQRVDNDXDFMSʥ$VVLVWDQW3URIHVVRU   7DNDVKL<DPDJXFKL3K'

6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU  7RP0DFSKHUVRQ3K'

2XUODERUDWRU\VWXGLHVQHXUDOFLUFXLWPHFKDQLVPVXQGHUO\LQJYDULRXVDGYDQFHGEUDLQIXQFWLRQVVXFKDVFRJQLWLYHOHDUQLQJDQGGHFLVLRQPDNLQJEHKDYLRUVXVLQJPROHFXODUWHFKQLTXHVIRUQHXUDOFLUFXLWVSHFLILFPDQLSXODWLRQ:HXVHVHYHUDOPRXVHPRGHOVWRUHYHDOPROHFXODUSDWKRORJLHVRIQHXURSV\FKLDWULFGLVHDVHV,QSDUWLFXODUZHIRFXVRQPROHFXODUPHFKDQLVPVRIJHQHHQYLURQPHQWLQWHUDFWLRQLQWKHSDWKRJHQHVLVRI

PHQWDOGLVRUGHUV:HDOVRSURPRWHWUDQVODWLRQDOUHVHDUFKIRUWDUJHWLQJPHQWDOGLVRUGHUVLQFROODERUDWLRQZLWKFOLQLFDOGHSDUWPHQWVDQGSKDUPDFHXWLFDOFRPSDQLHV

5HVHDUFK3URJUDPVʥ $QDO\VLVRIQHXUDOFLUFXLWPHFKDQLVPVLQDGYDQFHGEUDLQIXQFWLRQV
ʥ 0ROHFXODUDQDO\VLVRIQHXURSV\FKLDWULFSDWKRORJLHV
ʥ 7UDQVODWLRQDOUHVHDUFKRIPHQWDOGLVRUGHUV

5HVHDUFK7RSLFV‘ %DVDOJDQJOLDDUHWKHQHXUDOQXFOHLORFDWHGLQWKHFHQWHURIWKH㻌EUDLQ WKH\ DUH FRYHUHG DQG WDUJHWHG E\ WKH FHUHEUDO FRUWH[ 7KH
FRUWLFREDVDOJDQJOLDFLUFXLWLVHVVHQWLDOIRUPRWRUFRQWURODVZHOODV
DGYDQFHG EUDLQ IXQFWLRQ VXFK DV OHDUQLQJ FRJQLWLYH VRFLDO DQG
PHQWDO IXQFWLRQV 7KHUH DUH WZR GLVWLQFW SDWKZD\V LQ WKH EDVDOJDQJOLD FLUFXLW LH WKH GLUHFW DQG LQGLUHFW SDWKZD\V IURP WKH
VWULDWXPQXFOHXV DFFXPEHQV WR WKH VXEVWDQWLD QLJUD SDUVUHWLFXODWD+RZHYHUWKHLUUROHVLQDGYDQFHGEUDLQIXQFWLRQVUHPDLQ㻌XQFOHDU :H KDYH GHYHORSHG SDWKZD\VSHFLILF UHYHUVLEOH
QHXURWUDQVPLVVLRQ EORFNLQJ E\ VSHFLILFDOO\ DQG UHYHUVLEO\
H[SUHVVLQJWHWDQXVWR[LQZLWKLQQHXURQVLQWKHGLUHFWDQGLQGLUHFWSDWKZD\V +LNLGD HW DO 1HXURQ   :H KDYH IRXQG WKDW WKH
GLUHFWSDWKZD\LVLPSRUWDQWIRUUHZDUGOHDUQLQJDQGGUXJDGGLFWLRQZKLOHWKHLQGLUHFWSDWKZD\LVLPSRUWDQWIRUDYHUVLYHOHDUQLQJ +LNLGD
HW DO 1HXURQ   DQG WKDW GRSDPLQHUJLF LQSXW DQG SDWKZD\VSHFLILF3.$F$03VLJQDOLQJDUHFULWLFDOIRUOHDUQLQJ +LNLGDHWDO
31$6  <DPDJXFKL HW DO 31$6   :H DOVR IRXQG WKDWGRSDPLQH '/ UHFHSWRU VLJQDOLQJ LQ WKH LQGLUHFW SDWKZD\ LV
LPSRUWDQW IRU EHKDYLRUDO IOH[LELOLW\ LQ WKH UHZDUG OHDUQLQJ WDVNV LQ
WKH ,QWHOOL&DJH 0DFSKHUVRQ HW DO /HDUQ 0HP   DQG WKDWGRSDPLQH '/ UHFHSWRU LV UHTXLUHG IRU YLVXDO GLVFULPLQDWLRQ DQG
UHYHUVDOOHDUQLQJLQWRXFKVFUHHQFRJQLWLYHOHDUQLQJWDVNV 0RULWDHW
DO0RO1HXURSV\FKLDWU\ ◦ 30 ◦◦◦ Division of Integrated Protein Functions

8Sʁ%DVDO JDQJOLDFLUFXLW/HIW ʁWRXFKVFUHHQFRJQLWLYH OHDUQLQJWDVNVIRUPLFH

Institute for Protein Research Osaka University㻌㻌㻌‘ $QLPDO PRGHOVDUH UHTXLUHG IRU WUDQVODWLRQDOUHVHDUFK :H KDYH
㻌GHYHORSHG D PRXVH PRGHO IRU PHQWDO GLVRUGHUV WUDQVJHQLF PLFH

RYHUH[SUHVVLQJSDWLHQWW\SHPXWDQW',6&LQWKHIRUHEUDLQ +LNLGDHW㻌DO31$61LZDHWDO6FLHQFH 7KHVHJHQHWLFPRGHOPLFH㻌
VKRZ EHKDYLRUDO DEQRUPDOLWLHV DQG FKDQJHV LQ FRUWLFDO GRSDPLQH
㻌VLJQDOV DIWHU DGROHVFHQW VRFLDO LVRODWLRQ LQGLFDWLQJ JHQHWLF

㻌HQYLURQPHQWDOLQWHUDFWLRQVLQWKHSDWKRJHQHVLVRIPHQWDOGLVRUGHUV㻌  :HKDYHH[DPLQHGFRJQLWLYHIXQFWLRQVRIWKLVPRXVHPRGHOXVLQJWKHSODFHSUHIHUHQFHWDVN )LJ$ :HKDYHIRXQGWKDWWKHPXWDQW㻌',6& WUDQVJHQLF PLFH DUH LPSDLUHG LQ SODFH OHDUQLQJ DIWHU㻌

DGROHVFHQWVRFLDOLVRODWLRQ$VWKHSODFHFHOOLQWKHKLSSRFDPSDO&$

LVNQRZQWREHLPSRUWDQWIRUSODFHOHDUQLQJZHQH[WH[DPLQHGSODFHFHOODFWLYLW\E\ LQYLYRHOHFWURSK\VLRORJLFDOUHFRUGLQJ7KHUHZDVQRVLJQLILFDQW FKDQJH LQ WKH SODFH FHOO SURSHUW\ RI WKH PXWDQW ',6&

WUDQVJHQLF PLFH +RZHYHU UHZDUGUHODWHG DFWLYLW\ LQ WKH JRDO ]RQHGLVDSSHDUHGLQWKHSODFHFHOOVRIWKHPXWDQW',6&WUDQVJHQLFPLFH )LJ%  +D\DVKLHWDO1HXURVFL5HV LQGLFDWLQJKLSSRFDPSDOFLUFXLWDEQRUPDOLW\ .LWDQLVKLHWDO-36 






$ 7KH WHVWLQJ ER[ IRU WKH SODFH SUHIHUHQFH
WDVN$QLPDOVZHUHUHTXLUHGWRVWD\DWOHDVWVHFRQG LQ WKH XQPDUNHG JRDO ]RQH GDVKHGVTXDUH  DIWHU ZKLFK WLPH D IRRG SHOOHW ZDVGURSSHGIURPWKHRYHUKHDGGLVSHQVHU %([DPSOHSODFHILHOGPDSZLWKSXWDWLYHJRDO]RQH DFWLYLW\ /HIW  DQG ZLWKRXW JRDO]RQH
DFWLYLW\ 5LJKW 5HIV 

3XEOLFDWLRQV㻌 1HWZRUNPHFKDQLVPVRIKLSSRFDPSDOODWHUDOLW\SODFHFRGLQJDQGJRDOGLUHFWHGQDYLJDWLRQ.LWDQLVKLHWDO  㻌-3K\VLRO6FL

㻌

'RSDPLQH '/ UHFHSWRU LV UHTXLUHG IRU YLVXDO GLVFULPLQDWLRQ DQG UHYHUVDO OHDUQLQJ 0RULWD HW DO   0RO



1XFOHXV DFFXPEHQV GRSDPLQH 'UHFHSWRU H[SUHVVLQJ QHXURQV FRQWURO EHKDYLRUDO IOH[LELOLW\ LQ D SODFH

1HXURSV\FKLDWU\GLVFULPLQDWLRQWDVNLQWKH,QWHOOL&DJH0DFSKHUVRQHWDO  /HDUQ0HP

,PSDLUHGKLSSRFDPSDODFWLYLW\DWWKHJRDO]RQHRQWKHSODFHSUHIHUHQFHWDVNLQD',6&PRXVHPRGHO+D\DVKL

5ROH RI 3.$ VLJQDOLQJ LQ ' UHFHSWRUH[SUHVVLQJ QHXURQV LQ WKH FRUH RI WKH QXFOHXV DFFXPEHQV LQ DYHUVLYH

$GROHVFHQW VWUHVVLQGXFHG HSLJHQHWLF FRQWURO RI GRSDPLQHUJLF QHXURQV YLD JOXFRFRUWLFRLGV 1LZD HW DO  'LVWLQFWUROHVRIVHJUHJDWHGWUDQVPLVVLRQRIWKHVHSWRKDEHQXODUSDWKZD\LQDQ[LHW\DQGIHDU<DPDJXFKLHWDO



HWDO  1HXURVFL5HV

OHDUQLQJ<DPDJXFKLHWDO  3URF1DWO$FDG6FL86$

6FLHQFH

 1HXURQ

3DWKZD\VSHFLILF PRGXODWLRQ RI QXFOHXV DFFXPEHQVLQ UHZDUGDQG DYHUVLYH EHKDYLRU YLDVHOHFWLYHWUDQVPLWWHU

UHFHSWRUV+LNLGDHWDO  3URF1DWO$FDG6FL86$

'LVWLQFWUROHVRIV\QDSWLFWUDQVPLVVLRQLQGLUHFWDQGLQGLUHFWVWULDWDOSDWKZD\VWRUHZDUGDQGDYHUVLYHEHKDYLRU+LNLGDHWDO  1HXURQ 'RPLQDQWQHJDWLYH',6&WUDQVJHQLFPLFHGLVSOD\VFKL]RSKUHQLDDVVRFLDWHGSKHQRW\SHVGHWHFWHGE\PHDVXUHVWUDQVODWDEOHWRKXPDQV+LNLGDHWDO  3URF1DWO$FDG6FL86$

◦◦◦ 31 ◦

IPR
㻌

\
J
㻌/DERUDWRU\RI+RPHRVWDWLF,QWHJUDWLRQ㻌㻌㻌$VVRFLDWH3URIHVVRU

1REXDNL2NXPXUD3K'
QRNXPXUD#SURWHLQRVDNDXDFMS 

7KH /DERUDWRU\ RI +RPHRVWDWLF ,QWHJUDWLRQ LV HQJDJHG LQ VWXG\LQJ SURWHLQ DQG SHSWLGH PHWDEROLVP LQ

PDPPDOV:HKDYHSDUWLFXODUO\EHHQIRFXVLQJRQHQ]\PHVWKDWK\GURO\]HGLSHSWLGHVLQFOXGLQJKLVWLGLQHGLSHSWLGHVVXFKDVFDUQRVLQH5HFHQWO\ZHKDYHEHHQDSSO\LQJQDWLYHPDVVVSHFWURPHWU\WRDQDO\]HQRQFRYDOHQWSURWHLQFRPSOH[HVRIGLSHSWLGDVH&1WRHOXFLGDWHWKHPROHFXODUPHFKDQLVP RI LWVHQ]\PDWLF

UHDFWLRQ:HKDYHDOVREHHQSHUIRUPLQJGHYHORSPHQWDQGDSSOLFDWLRQRISURWHRPLFWHFKQLTXHVWRVWXG\

SURWHLQGHJUDGDWLRQDQGGLVHDVHELRPDUNHUV

5HVHDUFK3URJUDPV

0HWDEROLVPDQG)XQFWLRQRI+LVWLGLQH'LSHSWLGHV

3URWHRPLFDQDO\VLVRISURWHLQDQGSHSWLGHEUHDNGRZQ

3XEOLFDWLRQV

7KH]LQFIRUPRIFDUQRVLQHGLSHSWLGDVH &1 KDVGLSHSWLGDVHDFWLYLW\EXWLWVVXEVWUDWHVSHFLILFLW\LVGLIIHUHQW(YLGHQFH IRU DQ HVVHQWLDO UROH RI LQWUDGLPHU LQWHUDFWLRQ LQ FDWDO\WLFIXQFWLRQ RI FDUQRVLQH GLSHSWLGDVH ,, XVLQJ








IURPWKDWRIWKHPDQJDQHVHIRUP2NXPXUD1DQG7DNDR7  %%5&㻌

HOHFWURVSUD\LRQL]DWLRQPDVVVSHFWURPHWU\2NXPXUD17DPXUD-7DNDR7  3URWHLQ6FLHQFH&DUQRVLQHDQGWKHFRQWURORIEORRGJOXFRVH2NXPXUD1HWDO  LQ)RRGDQG1XWULWLRQDO&RPSRQHQWVLQ)RFXV56&SXEOLVKLQJ&DUQRVLQH GLSHSWLGDVH ,, 2NXPXUD 1   LQ +DQGERRN RI SURWHRO\WLF HQ]\PHV UG HG SS 

(OVHYLHU

'LYHUVLW\LQSURWHLQSURILOHVRILQGLYLGXDOFDOFLXPR[DODWHNLGQH\VWRQHV2NXPXUD1HWDO  3/RV2QH 

H

,GHQWLILFDWLRQRIFDUJRSURWHLQVVSHFLILFIRULPSRUWLQEHWDZLWKLPSRUWLQDOSKDDSSO\LQJDVWDEOHLVRWRSHODEHOLQJ

E\DPLQRDFLGVLQFHOOFXOWXUH 6,/$& EDVHGLQYLWURWUDQVSRUWV\VWHP.LPXUD0HWDO  -%LRO&KHP



6WUXFWXUDOEDVLVIRUVXEVWUDWHUHFRJQLWLRQDQGK\GURO\VLVE\PRXVHFDUQRVLQDVH&18QQRHWDO  -%LRO

&KHP

◦ 32 ◦◦◦ Division of Integrated Protein Functions

Division of Multiscale
Integrative Protein
ScienceLaboratory of Nuclear Network
Laboratory of Multiscale Structural Biology
Laboratory for Infection Systems
Laboratory of Visiting Scientists

IPR㻌

/DERUDWRU\ RI 1XFOHDU 1HWZRUN
㻌/DERUDWRU\RI1XFOHDU1HWZRUN㻌㻌㻌,QGHSHQGHQW$VVRFLDWH3URIHVVRU-XQNR.DQRK3K'
MNDQRK#SURWHLQRVDNDXDFMS '1$DVVRFLDWHVZLWKYDULRXVPDWHULDOVVXFKDVKLVWRQHVWRIRUPDFKURPRVRPH(XNDU\RWLF FHOOV FRQWDLQ OLQHDU FKURPRVRPHV WKH HQGV RI ZKLFK DUH FDOOHG WHORPHUHV 7HORPHUHV SOD\LPSRUWDQW UROHV LQ OLIH PDLQWHQDQFH VXFK DV FKURPRVRPH LQWHJULW\ UHJXODWLRQ RI FHOOXODU OLIHVSDQ DQG
VHPLSHUPDQHQWVXFFHVVLRQRIJHUPFHOOV:HDUHWU\LQJWRGLVFRYHUQRYHOIXQFWLRQVRIWHORPHUHIRFXVLQJ

RQWKH5DSSURWHLQZKLFKLVWKHFHQWHURIWKHWHORPHUHELQGLQJFRPSOH[LQILVVLRQ\HDVW:HZLOODOVRHOXFLGDWHZK\HXNDU\RWHVSUHIHUOLQHDUFKURPRVRPHV

7KH WHORPHUHDGMDFHQW FKURPRVRPDO GRPDLQ LV FDOOHG VXEWHORPHUH 6XEWHORPHUHV DUH GHYRLG RI D

WHORPHUH UHSHDW '1$VHTXHQFH EXW VKDUH KLJKO\KRPRORJRXV '1$ VHTXHQFHV DPRQJ WKHPVHOYHV 7KHSK\VLRORJLFDO IXQFWLRQV RI VXEWHORPHUHV KRZHYHU UHPDLQ ODUJHO\ XQFOHDU :H KDYH GHPRQVWUDWHG WKDWKLVWRQH+.PHPHGLDWHGKHWHURFKURPDWLQLVIRUPHGDWWKHVXEWHORPHULFKRPRORJRXVUHJLRQDQGWKDW

6JRDVKXJRVKLQSURWHLQLVORFDOL]HGDWWKHRWKHUUHJLRQRIWKHVXEWHORPHUHWRIRUPDKLJKO\FRQGHQVHGFKURPDWLQ VWUXFWXUH DQG FRQWULEXWHV WR WKHORFDO WUDQVFULSWLRQDO UHSUHVVLRQ DQG WKHPDLQWHQDQFHRI'1$UHSOLFDWLRQWLPLQJ:HZLOOLQYHVWLJDWHSK\VLRORJLFDOUROHV

RI

WKH

VXEWHORPHUHV LQ ILVVLRQ \HDVW KXPDQV DQGJUHDW DSHV IRU OLIH PDLQWHQDQFH GLYHUVLW\ DQGHYROXWLRQ

5HVHDUFK3URJUDPV)XQFWLRQVRIWHORPHUHVDQGWKHPHFKDQLVPVRIWHORPHUHSURWHLQFRPSOH[IRUPDWLRQ0HFKDQLVPVRIFKURPDWLQIRUPDWLRQDQGIXQFWLRQVRIVXEWHORPHUHV

5ROHVRIWKHOLQHDUIRUPRIHXNDU\RWLFFKURPRVRPHV&RPSDUDWLYHVWXG\RIWKHFKURPRVRPHHQGVWUXFWXUHVRIKXPDQVDQGJUHDWDSHV

3XEOLFDWLRQV

6XEWHORPHUHVFRQVWLWXWHDVDIHJXDUGIRUJHQHH[SUHVVLRQDQGFKURPRVRPHKRPHRVWDVLV7DVKLUR61LVKLKDUD<

6KXJRVKLQIRUPVDVSHFLDOL]HGFKURPDWLQGRPDLQDWVXEWHORPHUHVWKDWUHJXODWHVWUDQVFULSWLRQDQGUHSOLFDWLRQ






.XJRX.2KWD..DQRK-  1XFOHLF$FLGV5HVWLPLQJ 7DVKLUR6 +DQGD 7 0DWVXGD$ %DQ 7 7DNLJDZD 7 0L\DVDWR. ,VKLL . .XJRX . 2KWD . +LUDRND <

0DVXNDWD+.DQRK-  1DWXUH&RPPXQ

,GHQWLILFDWLRQRIWKHIXQFWLRQDOGRPDLQVRIWKHWHORPHUHSURWHLQ5DSLQ6FKL]RVDFFKDURP\FHVSRPEH)XMLWD,
7DQDND..DQRK-  3/R621(H7HORPHUHQXFOHDUHQYHORSHGLVVRFLDWLRQSURPRWHGE\5DSSKRVSKRU\ODWLRQHQVXUHVIDLWKIXOFKURPRVRPH

VHJUHJDWLRQ)XMLWD,1LVKLKDUD<7DQDND.7VXMLL+&KLNDVKLJH<:DWDQDEH<6DLWR0,VKLNDZD)+LUDRND<.DQRK-  &XUU%LRO

$FRQVHUYHGPRWLIZLWKLQ5$3SOD\VGLYHUVLILHGUROHVLQWHORPHUHSURWHFWLRQDQGUHJXODWLRQLQGLIIHUHQW

RUJDQLVPV&KHQ<5DL5=KRX=5.DQRK-5LEH\UH&<DQJ<=KHQJ+'DPD\3:DQJ)7VXMLL++LUDRND<6KRUH'+X+<&KDQJ6/HL0  1DWXUH6WUXF0RO%LRO 

◦ 34 ◦◦◦ Division of Multiscale Integrative Protein Science

Institute for Protein Research Osaka University

㻌

ER R \ R
X VF H 6 XF X
R RJ\
㻌/DERUDWRU\RI0XOWLVFDOH6WUXFWXUDO%LRORJ\
3URIHVVRU&RQFXUUHQW 㻌㻌㻌&RQFXUUHQW 㻌 $VVRFLDWH3URIHVVRU㻌㻌㻌㻌
9LVLWLQJ$VVRFLDWH3URIHVVRU $VVLVWDQW3URIHVVRU-XQLFKL7DNDJL3K'  *HQML.XULVX3K'.HQML,ZDVDNL3K'  'DPLHQ+DOO3K' 

1DR\XNL0L\D]DNL3K'

7DNDR$ULPRUL3K'

6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU  *HUW-DQ%HNNHU3K'

n0XOWLVFDOH 6WUXFWXUDO %LRORJ\| LV D WHUP GHYLVHG WR UHSUHVHQW QHZ W\SH RI OLIH VFLHQFH UHVHDUFK WUHQG

DLPHGDWHOXFLGDWLQJPROHFXODU PHFKDQLVPVRIFRPSOH[ELRORJLFDOV\VWHPVXVLQJPXOWLGLVFLSOLQDU\DQG

LQWHJUDWLYH H[SHULPHQWDO DSSURDFKHV UDQJLQJ IURP DWRPLF UHVROXWLRQ VWUXFWXUH GHWHUPLQDWLRQ WR

PDFURVFRSLF OLYH LPDJLQJ RI DQLPDOV 5HVHDUFKHUV LQ WKLV JURXS FRPELQH D ZLGH YDULHW\ RI PHWKRGVLQFOXGLQJFRPSXWHUVLPXODWLRQFU\RHOHFWURQPLFURVFRS\DQG105VSHFWURVFRS\WRDFFRPSOLVKWKLVDLP&RUUHODWLYH/LJKW(OHFWURQ0LFURVFRS\ &/(0

㻌7KH LPDJH UHQGHUHG XVLQJ 0ROPLO VKRZV D SDWKZD\FRQQHFWLQJELQGLQJFRQILJXUDWLRQVRIWKH&'.LQKLELWRU

&6SUHGLFWHGXVLQJIOH[LEOHPXOWLFDQRQLFDO0'GRFNLQJVLPXODWLRQVVWDUWLQJIURPWKHXQERXQGVWDWH OHIWVLGH 

7KH FRORUDORQJ WKH SDWKZD\ VKRZV WKH 30) REWDLQHG
YLDWKHUPRG\QDPLFLQWHJUDWLRQRIWKHIRUFHVVDPSOHGE\
FDQRQLFDO 0' VLPXODWLRQV VHHGHG IURP WKHVH ELQGLQJ
FRQILJXUDWLRQV %RWK WKH SUHGLFWHG VWDEOH ELQGLQJ
FRQILJXUDWLRQ &3. FRORU  DQG DIILQLW\ DJUHH YHU\ ZHOOZLWKH[SHULPHQWDOGDWD

7KHRUHWLFDODQGH[SHULPHQWDOPRGHOLQJRIDP\ORLGPLJUDWLRQWKURXJKDVL]HH[FOXVLRQFKURPDWRJUDSK\FROXPQ5HVHDUFK3URJUDPV

9LVXDOL]DWLRQRIELRPROHFXODUDUFKLWHFWXUHXVLQJLQWHJUDWLYHDSSURDFK&U\R(0EDVHGVWUXFWXUHGHWHUPLQDWLRQE\ODUJHVFDOHFRPSXWLQJDQGLPDJHDQDO\VLV(OXFLGDWLRQRIDP\ORLGIRUPDWLRQPHFKDQLVPE\FRPELQLQJH[SHULPHQWDODQGFRPSXWDWLRQDOPHWKRGV

3XEOLFDWLRQV$FFXUDWH SUHGLFWLRQ RI FRPSOH[ VWUXFWXUH DQG DIILQLW\IRU D IOH[LEOH SURWHLQ UHFHSWRU DQG LWV LQKLELWRU *HUW-DQ%HNNHU1DUXWRVKL.DPL\D0LWVXJX$UDNL,NXR)XNXGD<DVXVKL2NXQR+DUXNL1DNDPXUD  -&KHP7KHRU\&RPSXWt' K\EULG DQDO\VLV $SSURDFK IRU ELQGLQJ WKUHHGLPHQVLRQDO DWRPLF PRGHO E\ HOHFWURQ PLFURVFRS\ LPDJH$PXOWLSDWKZD\SHUVSHFWLYH RQSURWHLQDJJUHJDWLRQ,PSOLFDWLRQVIRUFRQWURORIWKHUDWHDQGH[WHQWRIDP\ORLGPDWFKLQJ0DWVXPRWR$HWDO  6FL5HS

IRUPDWLRQ'+DOO-.DUGRV+(GVNHV-$&DUYHU<*RWR  )(%6OHWWHUV

◦◦◦ 35 ◦IPR㻌/DERUDWRU\ IRU ,QIHFWLRQ 6\VWHPV㻌/DERUDWRU\IRU,QIHFWLRQ6\VWHPV㻌㻌㻌
6SHFLDOO\DSSRLQWHG
3URIHVVRU<XPLNR,PDL0' 3K'\LPDL#SURWHLQRVDNDXDFMS 

2XU ODERUDWRU\ DLPV WR XQGHUVWDQG KRVW UHVSRQVHV WR YLUXV LQIHFWLRQ DQG SDWKRJHQHVLV IRUPDWLRQ DV D

V\VWHP,QSDUWLFXODUZHDUHIRFXVLQJRQWKHUHVSRQVHVRIWKHKRVWQXFOHDUV\VWHPWRYLUDOLQIHFWLRQDQG

WKH PHFKDQLVP LQYROYHG LQWKH IRUPDWLRQ RI OHWKDO GLVHDVH VWDWHV :LWK WKH SURJUHVVRI UHFHQW JHQRPHDQDO\VLV WHFKQRORJ\ PDVV VSHFWURPHWU\ WHFKQRORJ\ ELRLQIRUPDWLFV DQDO\VLV WHFKQRORJ\ HWF LW KDV

EHFRPHSRVVLEOHWRDFTXLUHKXJHTXDQWLWDWLYHOLIHVFLHQFHGDWDRIPXOWLSOHOHYHOVHYHQLQWKHUHVHDUFKILHOGRI LQIHFWLRQ DQG LPPXQLW\ :KLOH FROODERUDWLQJ ZLWK GRPHVWLF DQG RYHUVHDV UHVHDUFK JURXSV ZH
FRPSUHKHQVLYHO\ DQDO\]H WKH TXDQWLWDWLYH OLIH VFLHQFH GDWD VXFK DV KRVW HSLJHQRPH SURILOLQJ P51$



WUDQVODWLRQSURILOHVPHWDERORPHDQDO\VLVVLQJOHFHOOWUDQVFULSWRPHDQDO\VLVKXPDQPLFURELRPHDQDO\VLVDQGHOXFLGDWHWKHQHWZRUNVLQYROYHGLQWKHPROHFXODUSDWKRORJ\DQGVHYHULW\RILQIHFWLRXVGLVHDVHV  %\

DSSO\LQJWKHJHQRPHV\QWKHVLVDQGJHQRPHHGLWLQJWHFKQRORJ\LQPRXVHPRGHOVZHFRQGXFWUHVHDUFK㻌DLPHGDWHVWDEOLVKLQJGUXJGLVFRYHU\GLDJQRVWLFPHWKRGVDQGSUHHPSWLYHPHGLFDOFDUHVEDVHGRQLW  

5HVHDUFK3URJUDPV '\QDPLFV RI KLJKHURUGHU FKURPDWLQ VWUXFWXUDO FKDQJHV WR YLUDO LQIHFWLRQ DQG WKHLU UROHV LQ GLVHDVHSDWKRJHQHVLV (OXFLGDWLRQRIKRVWDQGYLUXVP51$WUDQVODWLRQPHFKDQLVPWRYLUDOLQIHFWLRQ ,PPXQHQHXUDORUJDQLQWHUDFWLRQLQWKHSDWKRORJ\RIYLUXVLQIHFWLRQ

 3UHGLFWLRQRIG\QDPLFQHWZRUNUHVSRQVLEOHIRUWKHIRUPDWLRQRIVHYHUHSDWKRORJ\RIYLUXVLQIHFWLRQDQG
LWVDSSOLFDWLRQWRSUHHPSWLYHWKHUDS\

3XEOLFDWLRQV





<DPDJXFKL76X]XNL76DWR7HWDO7KH&&5127GHDGHQ\ODVHFRPSOH[FRQWUROV$WJGHSHQGHQWFHOOGHDWK

DQGKHDUWIXQFWLRQ6FL6LJQDO )HE  %ODQN7'HWMH&16SLH$HWDO3ULQ]0%UDLQ(QGRWKHOLDODQG(SLWKHOLDO6SHFLILF,QWHUIHURQ5HFHSWRU&KDLQ
'ULYHV9LUXV,QGXFHG6LFNQHVV%HKDYLRUDQG&RJQLWLYH,PSDLUPHQW,PPXQLW\ $SU  .DWDKLUD-'LPLWURYD/,PDL<HWDO17)OLNHGRPDLQRI7DSSOD\VDFULWLFDOUROHLQFDUJRP51$UHFRJQLWLRQ

DQGH[SRUW1XFOHLF$FLGV5HV)HE  

0RULWD 0 .XED . ,FKLNDZD $ HW DO 7KH OLSLG PHGLDWRU SURWHFWLQ ' LQKLELWV LQIOXHQ]D YLUXV UHSOLFDWLRQ DQG
LPSURYHVVHYHUHLQIOXHQ]D&HOO0DU  



,FKLNDZD $ .XED . 0RULWD 0 HW DO &;&/&;&5 HQKDQFHV WKH GHYHORSPHQW RI QHXWURSKLOPHGLDWHG

1HHO\*.XED.&DPPDUDWR$HWDO$JOREDOLQYLYR'URVRSKLOD51$LVFUHHQLGHQWLILHV127DVDFRQVHUYHGIXOPLQDQWOXQJLQMXU\RIYLUDODQGQRQYLUDORULJLQ$P-5HVSLU&ULW&DUH0HG-DQ  UHJXODWRURIKHDUWIXQFWLRQ&HOO  

,PDL < .XED . 1HHO\ ** HW DO ,GHQWLILFDWLRQ RI R[LGDWLYH VWUHVV DQG WROO OLNH UHFHSWRU  VLJQDOLQJ DV D NH\

SDWKZD\RIDFXWHOXQJLQMXU\&HOO  



,PDL<.XED.5DR6HWDO$QJLRWHQVLQFRQYHUWLQJHQ]\PHSURWHFWVIURPVHYHUHDFXWHOXQJIDLOXUH1DWXUH .XED . ,PDL < 5DR 6 HW DO $ FUXFLDO UROH RI DQJLRWHQVLQ FRQYHUWLQJ HQ]\PH  $&(  LQ 6$56FRURQDYLUXV-XO  LQGXFHGOXQJLQMXU\1DW0HG$XJ  

◦ 36 ◦◦◦ Division of Multiscale Integrative Protein Science

Institute for Protein Research Osaka University

㻌\J/DERUDWRU\RI9LVLWLQJ6FLHQWLVWV‘9LVLWLQJ3URIHVVRU 7RVKLR<DPD]DNL'6FLWRVKLR\DPD]DNL#ULNHQMS:H DUH GHYHORSLQJ 105 PHWKRGVIRU SURWHLQ VDPSOHV LQ WZR H[WUHPH FRQGLWLRQV ,Q RQH FRQGLWLRQ WKH

SURWHLQLVIRUFHGWRFKDQJHWKHIROGLQJVWDWH7KHIROGLQJLVFRQWUROOHGE\OLJKWLOOXPLQDWLRQDQGLVREVHUYHG

E\VROXWLRQ105:HUHFHQWO\VXFFHHGHGLQDWRPZLVHREVHUYDWLRQRIPHWDVWDEOHVWDWHLQWKHIROGLQJSDWK,QWKHRWKHUFRQGLWLRQDSURWHLQLVFDSWXUHGLQDELOD\HUPHPEUDQHDQGLVREVHUYHGE\VROLGVWDWH105
7RJHWULGRIVLJQDOFURZGLQJZKLFKKDPSHUVEDFNERQHDVVLJQPHQWZHDUHLQWURGXFLQJKLJKHUPDJQHWLFILHOGVDQGVHOHFWLYHLVRWRSHODEHOLQJ

5HVHDUFK3URJUDPVʥ /LJKWV\QFKURQRXV105REVHUYDWLRQIRUSURWHLQVWUXFWXUHFRQYHUVLRQ
ʥ 6ROLGVWDWH105IRUFRQGHQVHGVWDWHRISURWHLQV
ʥ 6ROXWLRQ105RIQRQFRGLQJ51$V

3XEOLFDWLRQV

1DJDVKLPD7RVKLR8HGD.HLVXNH1LVKLPXUD&KLDNL<DPD]DNL7RVKLR$QDO&KHPS 

.HQMLUR+DVKL.HQ]R'HJXFKL7RVKLR<DPD]DNLDQG7DGDVKL6KLPL]X&KHPLVWU\/HWWHUVS ‘9LVLWLQJ3URIHVVRU ,VDR6XHWDNH3K'

VXHWDNH#NRVKLHQDFMS, DP LQYHVWLJDWLQJ WKH PROHFXODU PHFKDQLVPV RI HSLJHQHWLFV DQG WKH HIIHFW RI QXWULWLRQ RQ HSLJHQHWLFUHJXODWLRQ,WLVUHSRUWHGWKDWWZRPDMRUUHJXODWLRQVDUHNQRZQWRFRQWULEXWHWRHSLJHQHWLFV2QHLVKLVWRQH

PRGLILFDWLRQVDQGWKHRWKHULV'1$PRGLILFDWLRQV7ULPHWK\ODWLRQRQO\VLQHRQ+LVWRQH+QHJDWLYHO\UHJXODWHV JHQH H[SUHVVLRQ KRZHYHU KRZ WKH PRGLILFDWLRQ LV UHFRJQL]HG LV QRW IXOO\ XQGHUVWRRG ,Q

PDPPDOVWKUHHLVRIRUPVRIKHWHURFKURPDWLQSURWHLQ +3 VSHFLILFDOO\ELQGWRWKHPHWK\ODWHGO\VLQH,DP LQYHVWLJDWLQJ LVRIRUPVSHFLILF ELQGLQJ RI +3 '1$ PHWK\ODWLRQ SDWWHUQV LQ JHQRPH LV HVWDEOLVKHG

GXULQJHPEU\RJHQHVLVDQGPDLQWDLQHG,DPH[DPLQLQJWKHSURSHUWLHVRIWKHHQ]\PHVUHVSRQVLEOH

5HVHDUFK3URJUDPV

0ROHFXODUPHFKDQLVPVXQGHUO\LQJWKHUHFRJQLWLRQRIPRGLILHGKLVWRQHV

8QGHUVWDQGLQJRIELRFKHPLFDOSURSHUWLHVRIHQ]\PHVUHVSRQVLEOHIRU'1$PHWK\ODWLRQ(SLJHQHWLFUHJXODWLRQE\1XWULWLRQ

3XEOLFDWLRQV ,VKL\DPD61LVKL\DPD$6DHNL<0RULWVXJX.ʞDQG1DNDQLVKL00RO&HOO   0LVKLPD<%UXHFNQHU/7DNDKDVKL6ʞDQG6XHWDNH,)(%6-  )XNX]DZD67DNDKDVKL67DFKLEDQD.DQG6XHWDNH,%LRRUJ0HG&KHP  0LVKLPD<-D\DVLQJKH&'ʞDQG6XHWDNH,1XFO$FLGV5HV ◦◦◦ 37 ◦IPR‘
㻌 9LVLWLQJ$VVRFLDWH3URIHVVRU.HQML6XJDVH3K'5KHR105LVDQ105LQVWUXPHQWWKDWFDQEHXVHGWRDQDO\]HWKHHIIHFWRIDSSOLHGVKHDUIRUFHRQDVDPSOH
LQUHDOWLPHDWDWRPLFUHVROXWLRQ:HKDYHVHWXSD5KHR105V\VWHPIRUWKHVXSHUKLJKVHQVLWLYLW\

0+]1058WLOL]LQJLWVKLJKVHQVLWLYLW\ZHDLPWRDQDO\]HPLQRUVWDWHVRISURWHLQVWKDWDUHIRUPHGGXULQJ

ILEULOIRUPDWLRQSURFHVVHV$OVRXVLQJVROLGVWDWH105ZHZLOODQDO\]HWKHVWUXFWXUHRILQVROXEOHILEULOVWKDWDFFXPXODWHLQWKH105VDPSOHWXEHDIWHU5KHR105PHDVXUHPHQWV

5HVHDUFK3URJUDPV'HYHORSPHQWRIDVXSHUKLJKVHQVLWLYLW\5KHR105LQVWUXPHQW(VWDEOLVKPHQWRIDSLSHOLQHIRUVWUXFWXUDODQDO\VLVRIDP\ORLGRJHQLFSURWHLQV

3XEOLFDWLRQV:DOLQGDHWDO2YHUYLHZRIUHOD[DWLRQGLVSHUVLRQ105VSHFWURVFRS\WRVWXG\SURWHLQG\QDPLFVDQGSURWHLQOLJDQG,ZDNDZDHWDO5HDOWLPHREVHUYDWLRQRIWKHLQWHUDFWLRQEHWZHHQWKLRIODYLQ7DQGDQDP\ORLGSURWHLQE\XVLQJ

LQWHUDFWLRQV&XUU3URWRF3URWHLQ6FLH  

KLJKVHQVLWLYLW\5KHR105 ,QW-0RO6FL  <DQDND HW DO (OXFLGDWLRQ RI SRWHQWLDO VLWHV IRU DQWLERG\ HQJLQHHULQJ E\ IOXFWXDWLRQ HGLWLQJ 6FL 5HS  



0RULPRWRHWDO+LJKVHQVLWLYLW\5KHR105VSHFWURVFRS\IRUSURWHLQVWXGLHV$QDO&KHP   

◦ 38 ◦◦◦ Division of Multiscale Integrative Protein ScienceResearch Center forState-of-the-Art Functional
Protein Analysis

Laboratory of Protein Profiling and Functional Proteomics
Laboratory of Protein Synthesis and Expression
Laboratory of Supramolecular Crystallography
Laboratory of Advanced Protein Characterization
NMR Group
EM Group
Laboratory of Protein Databases
Open Space Laboratory for Advanced Protein ScienceInstitute for Protein Research Osaka University

5HVHDUFK&HQWHUIRU6WDWHRIWKH$UW)XQFWLRQDO3URWHLQ$QDO\VLV

7KH 5HVHDUFK &HQWHU IRU 6WDWHRIWKH$UW )XQFWLRQDO 3URWHLQ $QDO\VLV 5&6)3  ZDV HVWDEOLVKHG LQ 
LQKHULWLQJWKHVXFFHVVRILWVSUHGHFHVVRUWKH5HVHDUFK&HQWHUIRU)XQFWLRQDODQG6WUXFWXUDO3URWHRPLFV,Q
UHFRJQLWLRQRIWKHLPSRUWDQFHRIH[SDQGLQJWKHVFRSHRIUHVHDUFKRXWVLGHWKHILHOGRISURWHRPLFVWKHFHQWHUQRZDLPVWRDQVZHUYDULRXVVFLHQWLILFTXHVWLRQVE\LQFRUSRUDWLQJDIXOOUDQJHRIFXWWLQJHGJHWHFKQRORJLHVRI
SURWHLQDQDO\VLVDQGZRUOGFODVVDQDO\WLFDOLQVWUXPHQWV7KHFHQWHUFRQVLVWVRIILYHODERUDWRULHVDQGRQHRSHQVSDFH ODERUDWRU\ DQG FRYHUV IROORZLQJ UHVHDUFK DUHDVSURWHLQ SURILOLQJ DQG IXQFWLRQDO SURWHRPLFV SURWHLQV\QWKHVLV DQG H[SUHVVLRQ VXSUDPROHFXODU FU\VWDOORJUDSK\ SURWHLQ LQIRUPDWLFV DGYDQFH SURWHLQFKDUDFWHUL]DWLRQDQGGDWDEDVHGHYHORSPHQW

/DERI6XSUDPROHFXODU&U\VWDOORJUDSK\/DERI3URWHLQ3URILOLQJDQG
)XQFWLRQDO3URWHRPLFV/DERI $GYDQFHG3URWHLQ&KDUDFWHUL]DWLRQ

2SHQ6SDFH/DE/DERI 
IRU$GYDQFHG
3URWHLQ'DWDEDVHV
3URWHLQ6FLHQFH/DERI3URWHLQ6\QWKHVLVDQG
([SUHVVLRQResearch Center for State-of-the-Art Functional Protein Analysis ◦◦◦ 41 ◦

IPR
㻌

\
J
㻌/DERUDWRU\RI3URWHLQ3URILOLQJDQG)XQFWLRQDO3URWHRPLFV㻌㻌
3URIHVVRU7RVKLIXPL7DNDR'6FL
WDN#SURWHLQRVDNDXDFMS 

0DVVVSHFWURPHWU\ 06 LVDZHOODFFHSWHGWHFKQLTXHIRUWKHDQDO\VLVRIFKHPLFDOVWUXFWXUHVRIELRORJLFDOFRPSRXQGV)RUWKHODVWWKUHHGHFDGHVZHKDYHEHHQZRUNLQJWRGHYHORSPHWKRGVIRUGHWHUPLQLQJSULPDU\VWUXFWXUHVDQGSRVWWUDQVODWLRQDOPRGLILFDWLRQVRISURWHLQVXVLQJ06,QFRQMXQFWLRQZLWKDFFXPXODWLQJ

SURWHLQ DQG JHQH VHTXHQFH GDWDEDVHV ZH DUH XVLQJ VWDWHRIWKHDUW 06 IRU ODUJHVFDOH SURWHLQ

LGHQWLILFDWLRQZKLFKLVLQGLVSHQVDEOHIRUSURWHRPLFVUHVHDUFK:HDSSO\WKHIROORZLQJGHYHORSHGPHWKRGVWRWKHVWUXFWXUDODQDO\VLVRIPLFURTXDQWLWLHVRISHSWLGHVSURWHLQVDQGWKHLUUHODWHGVXEVWDQFHV:HKDYH

IRXQG VHYHUDO QRYHO SRVWWUDQVODWLRQDO PRGLILFDWLRQV VXFK DV IDUQHV\O PRLHW\ DW WKH &WHUPLQDO &\V

KHWHURJHQHRXV IDWW\ DFLGV DW WKH 1WHUPLQDO *O\ H JJOXWDP\O  O\VLQH LQ FRUH KLVWRQHVSKRVSKDWLG\OHWKDQRODPLQHOLQNHGWRWKH&WHUPLQDO*O\SDOPLWROHR\OPRLHW\DW6HUHWF

5HVHDUFK3URJUDPV'HYHORSPHQWRIFKHPLFDODQDO\WLFDOPHWKRGVDQGVRIWZDUHIRUDQDO\VLVRISURWHLQSULPDU\VWUXFWXUH

E\PDVVVSHFWURPHWU\

0DVVVSHFWURPHWULFDQDO\VLVRISRVWWUDQVODWLRQDOPRGLILFDWLRQV'HYHORSPHQWRIFKHPLFDODQGDQDO\WLFDOPHWKRGVIRUSURWHRPLFV

6WXG\RQIUDJPHQWDWLRQLQPDVVVSHFWURPHWU\RISHSWLGHVDQGFDUERK\GUDWHV+DUGZDUHGHYHORSPHQWIRUKLJKVHQVLWLYLW\DQGKLJKDFFXUDF\PDVVVSHFWURPHWU\

5HVHDUFK7RSLFV‘ 7KH:QWIDPLO\RIVHFUHWHGVLJQDOSURWHLQVSOD\VD
NH\ UROH LQ QXPHURXV DVSHFWV RI HPEU\RJHQHVLV DV
ZHOODVLQFDUFLQRJHQHVLV,QFROODERUDWLRQZLWK3URI6
7DNDGD 2ND]DNL,QVWLWXWHIRU,QWHJUDWLYH%LRVFLHQFHV 
ZH IRXQG WKDW PXULQH :QWD LV DF\ODWHG DW D
FRQVHUYHG VHULQH UHVLGXH 6HU  ZLWK SDOPLWROHLF
DFLGϸ&  ZKLFK ZDV HVVHQWLDO IRU :QW

VHFUHWLRQ7KHOLSLGZDVQRWDVVDWXUDWHGDVRULJLQDOO\WKRXJKW )XUWKHUPRUH LQ FROODERUDWLRQ ZLWK 3URI $.LNXFKL*UDGXDWH 6FKRRO RI 0HGLFLQH 2VDND

8QLYHUVLW\  ZH GHWHUPLQHG PXOWLSOH 1JO\FRV\ODWLRQW\SHV IRU VHYHUDO :QWIDPLO\ SURWHLQV ZKLFK FRXOGUHJXODWH WKH GLUHFWLRQ RI :QW VHFUHWLRQ LQ SRODUL]HG

Fig.1. Novel fatty acid modification (palmitoleoyl modification at
Ser209) of Wnt protein essential for Wnt secretion (Dev. Cell 2006).

HSLWKHOLDOFHOOV

◦ 42 ◦◦◦ Research Center for State-of-the-Art Functional Protein Analysis

Institute for Protein Research Osaka University㻌㻌

‘ 3URWHLQV RIWHQ VXIIHU IURP YDULRXV FKHPLFDO㻌PRGLILFDWLRQV LQ YLYR ЎPLFURJOREXOLQ ЎP  D

347 days (t1/2)

Asn42

㻌VXEXQLWRI0+&FODVV,FLUFXODWHVDVDIUHHIRUPLQO

㻌EORRG 6LQFH ЎP KDV $VQ*O\ VHTXHQFHV DW WZR

O
OH
OH O

㻌VLWHVLQWKHPROHFXOHZKLFKDUHDPHQDEOHWRD⾫Ў
UHDUUDQJHPHQW ZH LQYHVWLJDWHG WKHLU VWDELOLW\ LQ㻌VROXWLRQ$VDUHVXOWGHDPLGDWLRQLVRPHUL]DWLRQDW

N
H

E2-Microglobulin

㻌$VQDQG$VQZLWKKDOIOLYHVRIDQGGD\V

18
8

N
H
O

N
H

R

L-Asp
L
L-As
A
Asp
As

Asn17
7

R
H
N

18

H O
OH
O

O

L-isoAsp
L-is
L-iso
iso
oA
Asp
sp
sp

33 days (t1/2)

㻌RFFXUUHGUHVSHFWLYHO\7KHUHDUUDQJHPHQWDWWKHVH
㻌VLWHV FDXVHG VRPH VWUXFWXUDO FKDQJH RI ЎP ,Q

Fig. 2 Quantitative Analysis of Deamidation and Isomerization in ȕ-㻌IDFW WKH PRGLILHG ЎP VKRZHG KLJKHU DIILQLW\ WR

Microglobulin by 18O Labeling (Anal. Chem. 2012).

&XLRQZKLFKZDVSUREHGE\(6,06㻌㻌㻌

‘ ,Q FROODERUDWLRQ ZLWK 3URI < 2KVXPL 1DWLRQDO

㻌

㻌

Apg8

,QVWLWXWH IRU %DVLF %LRORJ\ FXUUHQWO\ +RQRUDU\㻌

3URIHVVRURI7RN\R,QVWLWXWHRI7HFKQRORJ\ ZHIRXQGO㻌DQRYHOOLSLGPRGLILFDWLRQDW WKH& WHUPLQXVRI$SJ

Gly- NH-CH2CH2-O-P-O- CH2
Apg8

㻌ZLWKDSKRVSKROLSLGSKRVSKDWLG\OHWKDQRODPLQH 3( 㻌7KLV QRYHO OLSLGDWLRQ ZDV HVVHQWLDO IRU WKHOH CHO - CO-R’
CH2O-CO-R

15,003(PE)GDXWRSKDJRVRPHIRUPDWLRQLQDXWRSKDJ\㻌

㻌
㻌
㻌

12000

13000

14000

15000

16000

17000

㻌



18000

m/z

Fig. 3 Ubiquitination-like system mediates novel protein lipidation. (Nature 2000).


3XEOLFDWLRQV㻌 1.
㻌:QWEDVVRFLDWHGH[RVRPHVSURPRWHFDQFHUFHOOPLJUDWLRQDQGSUROLIHUDWLRQ+DUDGD7<DPDPRWR+.LVKLGD6

.LVKLGD0$ZDGD&7DNDR7.LNXFKL$  &DQFHU6FLHQFH  

㻌

2.

)UHH7KLRORI7UDQVWK\UHWLQLQ+XPDQ3ODVPD0RVW$FFHVVLEOHWR0RGLILFDWLRQ2[LGDWLRQ3HUHLUD&'0LQDPLQR

3.

)XQFWLRQDO /LQN EHWZHHQ $WJPHGLDWHG0LWRSKDJ\ DQG 3KRVSKROLSLG 0HWK\ODWLRQ 6DNDNLEDUD . (L\DPD $

17DNDR7  $QDO&KHP  6X]XNL6:6DNRK1DNDWRJDZD02NXPXUD17DQL0+DVKLPRWR$.RQGR2NDPRWR1.RQGR.DNXWD&$VDL(.LULVDNR+1DNDWRJDZD+.XJH27DNDR72KVXPL<2NDPRWR.  (0%2-  

4.

,GHQWLILFDWLRQRIFDUJRSURWHLQVVSHFLILFIRULPSRUWLQЎZLWKLPSRUWLQЍDSSO\LQJDVWDEOHLVRWRSHODEHOLQJE\DPLQRDFLGVLQFHOOFXOWXUH 6,/$& EDVHGLQYLWURWUDQVSRUWV\VWHP.LPXUD02NXPXUD1.RVH67DNDR7

,PDPRWR1  -%LRO&KHP

5.

4XDQWLWDWLYH$QDO\VLVRI'HDPLGDWLRQDQG,VRPHUL]DWLRQLQE20LFURJOREXOLQE\2/DEHOLQJ)XNXGD07DNDR7

6.

$IILQLW\7UDS 3RO\DFU\ODPLGH *HO (OHFWURSKRUHVLV $ 1RYHO 0HWKRG RI &DSWXULQJ 6SHFLILF 3URWHLQV E\ (OHFWUR $QDO&KHP

7UDQVIHU$ZDGD&6DWR77DNDR7  $QDO&KHPt

7.

0RQRXQVDWXUDWHG)DWW\$FLG0RGLILFDWLRQRI:QW3URWHLQ,WV5ROHLQ:QW6HFUHWLRQ7DNDGD56DWRPL<.XUDWD

8.

8ELTXLWLQDWLRQOLNH V\VWHP PHGLDWHV QRYHO SURWHLQOLSLGDWLRQ ,FKLPXUD < .LULVDNR 7 7DNDR 7 6DWRPL <78HQR11RULRND6.RQGRK+7DNDR7DQG7DNDGD6  'HY&HOO

6KLPRQLVKL<,VKLKDUD10L]XVKLPD1,VKLL72KVXPL01RGD7DQG2KVXPL<  1DWXUH



◦◦◦ 43 ◦

IPR
㻌

\
\
S
㻌/DERUDWRU\RI3URWHLQ6\QWKHVLVDQG([SUHVVLRQ㻌

3URIHVVRU -XQLFKL7DNDJL3K'
WDNDJL#SURWHLQRVDNDXDFMSʥ$VVRFLDWH3URIHVVRU$VVLVWDQW3URIHVVRU
6SHF$SSRLQW$VVLVW3URI

.HQML,ZDVDNL3K'<X.LWDJR3K', 1DR\XNL0L\D]DNL3K'
7DNDR$ULPRUL3K'$NLKLUR.DZDPRWR3K'

+RZGRWKLQJVZRUN"7KLVLVWKHTXHVWLRQPRVWLIQRWDOOVFLHQWLVWVDUHHDJHUWRDQVZHU2XUSDVVLRQLVWRXQUDYHOWKH PHFKDQLVP RI IXQFWLRQ RI SURWHLQV LQ D OLYLQJ RUJDQLVP ZKHUH WKH\ ZRUN ZLWK UHPDUNDEOH SUHFLVLRQ DV VPDOOPROHFXODUPDFKLQHV7KLVODEFDQEHGHVFULEHGDVDVWUXFWXUDOELRORJ\ODEEXWRXUJRDOLVQRWWKHGHWHUPLQDWLRQRIWKHWKUHHGLPHQVLRQDOVWUXFWXUHRISURWHLQV5DWKHUZH),567DLPWRVROYHWKHVWUXFWXUH7+(1SHUIRUPELRFKHPLFDOELRSK\VLFDODQGFHOOELRORJLFDOH[SHULPHQWVWRGUDZQRYHOSLFWXUHVDERXWWKHPROHFXODUPHFKDQLVPRISURWHLQVWDNLQJDGYDQWDJHRIWKHVWUXFWXUDOLQIRUPDWLRQWKDWLVH[FOXVLYHWRXV  :HDUHPRVWO\IRFXVHGRQWKHPROHFXODULQWHUDFWLRQVEHWZHHQFHOOVXUIDFHUHFHSWRUVDQGWKHLUH[WUDFHOOXODUOLJDQGVLPSOLFDWHGLQVLJQDOWUDQVGXFWLRQLQDZLGHYDULHW\RIELRORJLFDOFRQWH[WVUDQJLQJIURPGHYHORSPHQWQHXURELRORJ\DQGLPPXQLW\2XUVWXG\IRFXVHVRQTXHVWLRQVVXFKDVKRZUHFHSWRUVUHFRJQL]HWKHLUVSHFLILFOLJDQGVKRZWKLVUHFRJQLWLRQOHDGVWRVWUXFWXUDOFKDQJHLQWKHUHFHSWRUFRPSOH[DQGKRZWKHLQIRUPDWLRQFURVVHVWKHSODVPDPHPEUDQHZLWKRXWWUDQVSRUWLQJDFKHPLFDOHQWLW\ 
2XU DSSURDFK LV PXOWLIDFHWHG $V WKH PHWKRGRORJ\ IRU VWUXFWXUDO DQDO\VLV ZH XWLOL]H ;UD\ FU\VWDOORJUDSK\ ZKLFKGHWHUPLQHV'VWUXFWXUHRISURWHLQVDWDWRPLFUHVROXWLRQDQGWUDQVPLVVLRQHOHFWURQPLFURVFRS\ 7(0 ZKLFKFDQ
GHULYHWKHVWUXFWXUHRISURWHLQFRPSOH[HVWRRODUJHIRU;5'RUYLVXDOL]HWKHVKDSHRISURWHLQVLQWKHLUWUXHELRORJLFDOHQYLURQPHQW HJ ZLWKLQ FHOOV  7KH ODWWHU H[SHUWLVH LQFOXGHV FXWWLQJHGJH WHFKQRORJLHV VXFK DV FU\RHOHFWURQPLFURVFRS\DQGHOHFWURQWRPRJUDSK\,QRUGHUWREDFNWKHVHVWUXFWXUDOHIIRUWVZHKDYHDOVRGHYHORSHGDQDUUD\RILQ
KRXVHWHFKQRORJLHVFULWLFDOIRUWKHSURGXFWLRQRIKLJKTXDOLW\UHFRPELQDQWSURWHLQVXVLQJPDPPDOLDQFHOOH[SUHVVLRQV\VWHP

5HVHDUFK3URJUDPV
㻌


㻌
㻌

㻌

6WUXFWXUH DQG IXQFWLRQ RI H[WUDFHOOXODU OLJDQGV DQG WKHLU UHFHSWRUV LPSOLFDWHG LQ FHOO DGKHVLRQ DQG QHXUDOJXLGDQFHPRUSKRJHQHVLV
6WUXFWXUHJXLGHGPROHFXODUGHVLJQRIQRYHOSURWHLQV
&RUUHODWLYHVWUXFWXUDODQDO\VLVE\PXOWLGLVFLSOLQDU\DSSURDFK'HYHORSPHQWRIKLJKTXDOLW\UHFRPELQDQWSURWHLQSURGXFWLRQV\VWHP
'YLVXDOL]DWLRQRIFRQIRUPDWLRQDOO\KHWHURJHQHRXVPDFURPROHFXOHVXVLQJHOHFWURQPLFURVFRS\

5HVHDUFK7RSLF‘ 6HPDSKRULQVDUHDFODVVRID[RQJXLGDQFHPROHFXOHVDQGELQGWRFHOOVXUIDFHUHFHSWRUSOH[LQV6HPDSKRULQSOH[LQ

VLJQDOLQJLVLPSOLFDWHGQRWRQO\LQQHXURQDOFHOOVEXWDOVRLQLPPXQHFHOOVERQHFHOOVDQGFDQFHUFHOOVPDNLQJLWDQ
LQWHQVLYH WDUJHW RI GUXJ GHYHORSPHQW ,Q  ZH UHSRUWHG WKH ILUVW FU\VWDO VWUXFWXUH RI WKH VHPDSKRULQSOH[LQ
VLJQDOLQJ FRPSOH[ UHI   6LQFH WKHQ ZH KDYH EHHQ DFWLYHO\ HQJDJHG LQ UHVHDUFKHV DLPHG DW XQUDYHOLQJ WKH
PHFKDQLVPRIVHPDSKRULQSOH[LQVLJQDOLQJXVLQJERWKFU\VWDOORJUDSK\DQGHOHFWURQPLFURVFRS\ ,QDGGLWLRQWR WKH
VWUXFWXUDODQDO\VHVZHWU\WRFODULI\WKHVWUXFWXUHIXQFWLRQUHODWLRQVKLSDERXWWKLVLPSRUWDQWVLJQDOLQJSDWKZD\XVLQJFHOOEDVHGDVVD\VHQDEOLQJXVWRGLVFRYHUQRYHOFODVVRILQKLELWRUWKDWFDQSRWHQWLDOO\EHFRPHDGUXJOHDG UHI 

1HJDWLYHVWDLQ(0LPDJH OHIW 
DQGLWVH[SHFWHGVWUXFWXUDOPRGHOULJKW RISOH[LQHFWRGRPDLQ&U\VWDOVWUXFWXUHRI
VHPDSKRULQSOH[LQFRPSOH[◦ 44 ◦◦◦ Research Center for State-of-the-Art Functional Protein Analysis&HOOPRUSKRORJ\EHIRUH OHIW DQGDIWHUULJKW   VWLPXODWLRQE\VHPDSKRULQInstitute for Protein Research Osaka University㻌஁പ࣯ߑ஁പ࣯ߑଆਫ਼෼ָ‫ڂݜ‬෨໵
஁പ
࣯ߑଆਫ਼
ଆਫ਼෼ָ
෼ָ‫ڂݜ‬
෼ָ‫ڂݜ‬෨໵‫ڂݜ‬෨໵㻌ଆਫ਼㻌 ‘ &HOO DGKHVLRQ PROHFXOHV VXFK DV LQWHJULQV DQGFDGKHULQVSOD\IXQGDPHQWDOUROHVLQWKHGHYHORSPHQWDVZHOO DV RUJDQRJHQHVLV RI PHWD]RDQ VSHFLHV LQFOXGLQJ㻌 KXPDQV DQG WKHLU GHIHFWV OHDG WR YDULRXV GLVHDVHVLQFOXGLQJ FDQFHU :H KDYH GHWHUPLQHG WKH FU\VWDO㻌VWUXFWXUHRILQWHJULQDEWKHPDMRUFHOOVXUIDFHUHFHSWRU㻌 FRPSOH[ IRU H[WUDFHOOXODU PDWUL[ LQ PDPPDOV DQGFODULILHG LWV OLJDQG UHFRJQLWLRQ PHFKDQLVP DW DWRPLF
㻌UHVROXWLRQ UHI $OVRZHKDYHVXFFHHGHGLQYLVXDOL]LQJ㻌 WKH ' DUFKLWHFWXUH RI JDQJDQWLF FDGKHULQ E\
WUDQVPLVVLRQ HOHFWURQ PLFURVFRS\ UHI   ZKLFK ZDV㻌LPSRVVLEOH E\ FU\VWDOORJUDSK\ GXH WR WKH KLJKO\㻌 SRO\PRUSKLFQDWXUHRIWKHPROHFXOH㻌‘ $O]KHLPHUbVGLVHDVHLVDSUHYDOHQWQHXURGHJHQHUDWLYH㻌&U\VWDOVWUXFWXUHRILQWHJULQDE

7(0LPDJHRIJDQJDQWLF FDGKHULQ)$7 OHIW DQGLWVGRPDLQILWWLQJPRGHO ULJKW 㻌㻌GLVHDVHZLWKQRHVWDEOLVKHGWUHDWPHQWVDQGLVWKRXJKWWREHFDXVHGE\H[WUDFHOOXODUDFFXPXODWLRQRILQVROXEOH㻌
DJJUHJDWHVFRQWDLQLQJDP\ORLGE $E SHSWLGHLQEUDLQV㻌,QZHGLVFRYHUHG WKDW DQHXURQDOUHFHSWRUFDOOHG
VRU/$ FDQ FDSWXUH $EDQG WUDQVSRUW LW WR WKH O\VRVRPH㻌
IRUGHJUDGDWLRQUHGXFLQJWKHULVNRI$',QDVXEVHTXHQW㻌VWXG\ZHVROYHG WKHFU\VWDO VWUXFWXUHRIVRU/$9SVS 'HJUDGDWLRQRI$E
E\QHXURQDOVRU/$
GRPDLQDQGIRXQGWKDWWKLVGRPDLQVHUYHVDVDFDSWXULQJ㻌GHYLFH IRU $E SHSWLGH DQG SUREDEO\ RWKHU SRWHQWLDOO\㻌
DP\ORLGRJHQLFSDSWLGHDQGWKXVnJXDUGV|EUDLQVIURP$'
UHI 
㻌

&U\VWDOVWUXFWXUHRIVRU/$9SVSGRPDLQ㻌‘ :HGHYHORSHGDFXVWRPPDGHKLJKDIILQLW\HSLWRSHWDJJLQJV\VWHPFDOOHG3$

㻌WDJ ZKLFK HQDEOHV RQHVWHS FRPSOHWH SXULILFDWLRQ RI UHFRPELQDQW SURWHLQV
SUHVHQWLQDFUXGHVDPSOHDWORZDEXQGDQFH UHI 3$WDJZDVOLFHQVHGWRD㻌UHDJHQW FRPSDQ\ DQG VROG ZRUOGZLGH PDNLQJ LW DYDLODEOH WR WKH FRPPXQLW\㻌8VLQJ3$WDJDQGVHYHUDORWKHUWHFKQRORJLHVGHYHORSHGLQWKHODEZHVXFFHHGHG

LQSXULI\LQJKLJKTXDOLW\:QWSURWHLQVZKLFKKDVDKXJHLPSDFWRQWKHVWHPFHOO㻌ELRORJ\ DQG UHJHQHUDWLYH PHGLFLQH UHI   :H DOVR GHYHORSHG D QRYHO VPDOO㻌DQWLERG\GHVLJQn)YFODVS|DQGDSSOLHGLWWRFKDSHURQHDVVLVWHGFU\VWDOORJUDSK\RIKLJKYDOXHWDUJHWSURWHLQV UHI 㻌

6WUXFWXUHRIDQRYHOVPDOODQWLERG\IRUPDWn)YFODVS| 

3XEOLFDWLRQV㻌 'HYHORSPHQWRIDQHZSURWHLQODEHOLQJV\VWHPWRPDSVXEXQLWVDQGGRPDLQVRIPDFURPROHFXODUFRPSOH[HVIRU
HOHFWURQPLFURVFRS\%URZQHWDO  -6WUXFW%LRO 㻌  ,PPXQHHYDVLRQRI3ODVPRGLXPIDOFLSDUXPE\5,),1YLDLQKLELWRU\UHFHSWRUV6DLWRHWDO㻌 1DWXUH  )YFODVS$QDUWLILFLDOO\GHVLJQHGVPDOODQWLERG\IUDJPHQWZLWKLPSURYHGSURGXFWLRQFRPSDWLELOLW\VWDELOLW\DQG
FU\VWDOOL]DELOLW\$ULPRULHWDO  6WUXFWXUH  $OORVWHULFLQKLELWLRQRIDVHPDSKRULQ'UHFHSWRUSOH[LQ%E\DKLJKDIILQLW\PDFURF\FOLFSHSWLGH0DWVXQDJDHWDO
 &HOO&KHP%LRO

 7DLORUHGSODFHPHQWRIDWXUQIRUPLQJ3$WDJLQWRWKHVWUXFWXUHGGRPDLQRIDSURWHLQWRSUREHLWVFRQIRUPDWLRQDOVWDWH)XMLLHWDO  -&HOO6FLHQFH $FWLYH DQG ZDWHUVROXEOH IRUP RI OLSLGDWHG :QW SURWHLQ LV PDLQWDLQHG E\ D VHUXP JO\FRSURWHLQ DIDPLQЍ
DOEXPLQ0LKDUDHWDO  H/LIHH  6WUXFWXUDOEDVLVIRUDP\ORLGRJHQLFSHSWLGHUHFRJQLWLRQE\VRU/$.LWDJRHWDO  1DWXUH6WUXFW0RO%LRO


 *LDQWFDGKHULQV)DWDQG'DFKVRXVVHOIEHQGWRRUJDQL]HSURSHUO\VSDFHGLQWHUFHOOXODUMXQFWLRQV7VXNDVDNLHWDO
 3URF1DWO$FDG6FL86$

 &U\VWDOVWUXFWXUHRIЍЎLQWHJULQHFWRGRPDLQ$WRPLFGHWDLOVRIWKHILEURQHFWLQUHFHSWRU1DJDHHWDO  -&HOO%LRO 

 6WUXFWXUDOEDVLVIRUVHPDSKRULQVLJQDOLQJWKURXJKWKHSOH[LQUHFHSWRU1RJLHWDO  1DWXUH

◦◦◦ 45 ◦

IPR
㻌

\
S
\
J S \
㻌/DERUDWRU\RI6XSUDPROHFXODU&U\VWDOORJUDSK\㻌㻌㻌
3URIHVVRU$WVXVKL1DNDJDZD'6FLDWVXVKL#SURWHLQRVDNDXDFMS $VVRFLDWH3URIHVVRU      0DPRUX6X]XNL3K'$VVRFLDWH3URIHVVRU &RQFXUUHQW   (LNL<DPDVKLWD3K'

7KHUH DUH YDULRXV ELRORJLFDO PDFURPROHFXODU DVVHPEOLHV FRQVLVWLQJ RI SURWHLQV QXFOHLF DFLGV VXJDUVOLSLGVDQGRWKHUVXEVWDQFHVLQOLYLQJFHOOV7KHVHPDFURPROHFXODUDVVHPEOLHVSOD\NH\UROHVLQDOOOLYLQJV\VWHPV 2XU ODERUDWRU\ ZRUNVRQ VWUXFWXUH GHWHUPLQDWLRQ RI ELRORJLFDO PDFURPROHFXODU DVVHPEOLHV DV
ZHOO DV SURWHLQV ZKLFK SOD\ LPSRUWDQW UROHV LQ ELRORJLFDO V\VWHPV XVLQJ ;UD\ FU\VWDOORJUDSK\'HYHORSPHQWRIWRROVIRU;UD\FU\VWDOVWUXFWXUHGHWHUPLQDWLRQRIELRORJLFDOPDFURPROHFXODUDVVHPEOLHVLQFOXGLQJV\QFKURWURQUDGLDWLRQEHDPWLPHDW63ULQJLVDOVRRQHRIRXUPDLQZRUNV

5HVHDUFK3URJUDPVʥ ;UD\VWUXFWXUHGHWHUPLQDWLRQRIPDFURPROHFXODUDVVHPEOLHVDQGSURWHLQV
ʥ 'HYHORSPHQW PHWKRGRORJLHV IRU ;UD\ VWUXFWXUH GHWHUPLQDWLRQ RI ELRORJLFDO PDFURPROHFXODU
DVVHPEOLHVXVLQJV\QFKURWURQUDGLDWLRQDQG;UD\IUHHHOHFWURQODVHU
ʥ 'HYHORSPHQWRIGDWDSURFHVVLQJDOJRULWKPRIGLIIUDFWLRQGDWDIURPPLFURFU\VWDOV

5HVHDUFK7RSLFV㻌‘ 3URWHLQ FU\VWDOORJUDSK\ KDV PDGH JUHDW SURJUHVV LQ UHFHQW \HDUVDQGPDQ\FRPSOH[SURWHLQVWUXFWXUHVKDYHEHHQGHWHUPLQHGGXULQJWKLV㻌

WLPH +RZHYHU WKHUH DUH VWLOO PDQ\ EDUULHUV WR EH RYHUFRPH LQ WKH

㻌SURFHVV RI
㻌 VWUXFWXUH GHWHUPLQDWLRQ +LJK TXDOLW\ GLIIUDFWLRQ GDWD

FROOHFWLRQLVRQHRIWKHPRVWLPSRUWDQWVWHSVIRUVXFFHVVRIVWUXFWXUH
GHWHUPLQDWLRQ )RU WKLV SXUSRVH ZH DUH PDQDJLQJ WKH V\QFKURWURQUDGLDWLRQ EHDPOLQH DW 63ULQJ IRU KLJK TXDOLW\ GLIIUDFWLRQ GDWD
FROOHFWLRQ IURP ELRORJLFDO PDFURPROHFXODU DVVHPEO\ FU\VWDOV 7KH
EHDPOLQH %/;8 LV QDPHG WKH EHDPOLQH IRU ELRORJLFDO㻌PDFURPROHFXODUDVVHPEOLHVRU,35EHDPOLQH 7DQSDNXNHQEHDPOLQH 
7KLVEHDPOLQHDLPVWRFROOHFWKLJKTXDOLW\GLIIUDFWLRQGDWDIURPVPDOO%/;8DW63ULQJVL]HDQGODUJHXQLWFHOOFU\VWDOVVXFKDVPHPEUDQHSURWHLQFRPSOH[HV㻌

SURWHLQ FRPSOH[HV SURWHLQ DQG QXFOHLF DFLG FRPSOH[HV DQG YLUXVHV㻌'DWDFROOHFWLRQIURPRUGLQDU\SURWHLQFU\VWDOVDOVRFDQEHDFKLHYHG
㻌7KLVEHDPOLQHLVDYDLODEOHWRDQ\VFLHQWLVWLQWKHZRUOGDVRQHRIWKHSURJUDPVRIWKH-RLQW8VDJHDQG5HVHDUFKIRU
㻌3URWHLQV,QWHUQDWLRQDOFROODERUDWLRQEHWZHHQWKH1DWLRQDO6\QFKURWURQ5DGLDWLRQ5HVHDUFK&HQWHU 1655& LQ7DLZDQLVVWLPXODWLQJWKHDFWLYLWLHVRIWKHEHDPOLQH

◦ 46 ◦◦◦ Research Center for State-of-the-Art Functional Protein Analysis

Institute for Protein Research Osaka University㻌㻌‘ (OHFWULFVLJQDOVDUHLPSRUWDQWLQPDQ\OLYLQJSURFHVVHVVXFKDVQHUYH㻌VLJQDO WUDQVPLVVLRQ DQG KHDUW EHDW 5HFHQWO\ QHZ YROWDJHVHQVLQJ㻌SURWHLQVKDYHEHHQLGHQWLILHGDQG WKHVHPROHFXOHVWUDQVGXFHHOHFWULFDO㻌VLJQDOV LQWR LQWUDFHOOXODU VLJQDOV LQ D GLIIHUHQW ZD\ IURP WKH FODVVLFDO㻌YROWDJHJDWHG LRQ FKDQQHOV 2XU SURMHFW DLPV WR UHYHDO WKH G\QDPLF
㻌PHFKDQLVP RI VLJQDO WUDQVGXFWLRQ E\ WKH QHZ YROWDJHVHQVLQJ SURWHLQVEDVHGRQWKHLUDWRPLFVWUXFWXUH7KHVHUHVXOWVZLOORSHQDQHZSDUDGLJP㻌RI OLIH VFLHQFH 2XU UHVXOWV ZLOO JLYH YDOXDEOH LQIRUPDWLRQ IRU VWUDWHJLF

9ROWDJHVHQVLQJSURWHLQIDPLO\㻌

GHYHORSPHQWRIQHZWRROVIRUYLVXDOL]DWLRQRIHOHFWULFDOVLJQDODQGZLOOEH㻌H[WHQGHGWRVWXGLHVIRUPHGLFDODSSOLFDWLRQ㻌㻌㻌㻌‘ :H DUH ZRUNLQJ RQ VWUXFWXUDO VWXGLHV RI ELRORJLFDO PDFURPROHFXODU㻌DVVHPEOLHVVXFKDVSODQUHRYLUXVHVFKORUHOODYLUXVDQGPXOWLGUXJHIIOX[SXPSFRPSOH[HV2QHRIRXUPDLQWDUJHWVLVD 5LFHGZDUIYLUXV 5'9 RIZKLFKDWRPLFVWUXFWXUHVRIFDSVLGKDVEHHQGHWHUPLQHG:HDUHZRUNLQJWR

GHWHUPLQHWKHFRPSOHWHVWUXFWXUHRIWKHYLUXVLQFOXGLQJWKHWUDQVFULSWLRQDOFRPSOH[JHQRPHDQGWKHSURWHLQFRPSRQHQWWKDWLVUHTXLUHGIRULQIHFWLRQ,Q DGGLWLRQ QRQVWUXFWXUH SURWHLQV ZKLFK DUH HVVHQWLDO IRU DVVHPEO\PXOWLSOLFDWLRQDQGLQIHFWLRQWRWKHQH[WFHOODUHDOVRWDUJHWHG:HDLPWRXQGHUVWDQGWKHOLIHF\FOHRI5'9IURPWKHDWRPLFVWUXFWXUHVRISURWHLQVRI

5'9

0RGHORIPXOWLSOLFDWLRQSURFHVVRI

5LFHGZDUIYLUXVLQDFHOO

:H DUH DOVR ZRUNLQJ RQ WKH FRKHUHQW ;UD\ GLIIUDFWLRQ LPDJLQJ &', RU
&;', RIDVLQJOHSDUWLFOHRIYLUXVXVLQJ;UD\IUHHHOHFWURQODVHU ;)(/ XQGHUWKHLQWHUQDWLRQDOFROODERUDWLRQSURJUDP

3XEOLFDWLRQV㻌 +LHUDUFKLFDO VWUXFWXUH DVVHPEO\ PRGHO RI ULFH GZDUI YLUXV SDUWLFOH IRUPDWLRQ 1DNDJDZD $ 0L\D]DNL 1 DQG㻌

+LJDVKLXUD$  %LRSK\VLFDO5HYLHZV

㻌

 6WUXFWXUDOEDVLVIRUWKHDVVHPEO\RIWKH5DJXODWRU5DJ*73DVHFRPSOH[<RQHKDUD51DGD61DNDL71DNDL
0.LWDPXUD$2JDZD$1DNDWVXPL+1DND\DPD.,/L66WDQGOH\'0<DPDVKLWD(1DNDJDZD$

DQG2NDGD0  1DWXUH&RPPXQLFDWLRQV   &U\VWDOVWUXFWXUHVRI2SU1DQG2SU-RXWHUPHPEUDQHIDFWRUVRIPXOWLGUXJWULSDUWLWHHIIOX[SXPSVRI3VHXGRPRQDVDHUXJLQRVD<RQHKDUD5<DPDVKLWD(1DNDJDZD$  3URWHLQV6WUXFWXUH)XQFWLRQDQG%LRLQIRUPDWLFV


 ;UD\ FU\VWDO VWUXFWXUH RI YROWDJHJDWHG SURWRQ FKDQQHO 7DNHVKLWD . 6DNDWD 6 <DPDVKLWD ( )XMLZDUD <.DZDQDEH $.XURNDZD72NRFKL<0DWVXGD01DULWD+2NDPXUD<DQG1DNDJDZD$  1DWXUH

6WUXFWXUDO 0ROHFXODU%LRORJ\

 $ QHZ SURWHLQ FRPSOH[ SURPRWLQJ WKH DVVHPEO\ RI 5DG ILODPHQWV 6DVDQXPD + 7DZDUDPRWR 06 /DR -

+RVDND+6DQGD(6X]XNL0<DPDVKLWD(6KLQRKDUD0+XQWHU11DNDJDZD$DQG6KLQRKDUD$  

1DWXUH&RPPXQLFDWLRQV

 6WUXFWXUDO LQVLJKW LQWR PDLQWHQDQFH PHWK\ODWLRQ E\ PRXVH '1$ PHWK\OWUDQVIHUDVH  'QPW  7DNHVKLWD .6XHWDNH,<DPDVKLWD(6XJD01DULWD+1DNDJDZD$DQG7DMLPD6  3URFHHGLQJVRIWKH1DWLRQDO

$FDGHP\RI6FLHQFHV86$ &U\VWDO 6WUXFWXUH RI WKH 0HPEUDQH )XVLRQ 3URWHLQ 0H[$ RI WKH 0XOWLGUXJ 7UDQVSRUWHU LQ 3VHXGRPRQDVDHUXJLQRVD $NDPD + 0DWVXXUD 7 .DVKLZDJL 6 <RQH\DPD + 1DULWD 6 7VXNLKDUD 7 1DNDJDZD $ DQG1DNDH7  -RXUQDORI%LRORJLFDO&KHPLVWU\◦◦◦ 47 ◦IPR㻌/DERUDWRU\RI$GYDQFHG3URWHLQ&KDUDFWHUL]DWLRQ
㻌/DERUDWRU\RI$GYDQFHG3URWHLQ&KDUDFWHUL]DWLRQ
7KLV ODERUDWRU\ LV ZRUNLQJ RQ WKH GHYHORSPHQW DQG DSSOLFDWLRQ RI VHYHUDO LPSRUWDQW PHWKRGRORJLHV IRUDGYDQFHGFKDUDFWHUL]DWLRQRISURWHLQPROHFXOHVDQGSOD\VDQLPSRUWDQWUROHLQWHFKQLFDOLQQRYDWLRQVIRU

SURWHLQDQDO\VHVDQGPHDVXUHPHQWV)DFXOW\PHPEHUVLQWKLVODERUDWRU\DUHRUJDQL]HGLQWRWKUHHUHVHDUFKJURXSV D JURXS XVLQJ 105 ZLWK VXSHU KLJK PDJQHW ILHOG D JURXS IRU ;UD\ FU\VWDOORJUDSK\ XVLQJV\QFKURWURQ UDGLDWLRQ DW 63ULQJ DQ HOHFWURQ PLFURVFRSLF LPDJLQJ JURXS DQG RQH WHFKQLFDO VXSSRUWJURXSIRUFKHPLFDODQDO\VHVRISURWHLQPROHFXOHV

‘6\QFKURWURQ5DGLDWLRQ5HVHDUFK*URXS
3URIHVVRU &RQFXUUHQW$VVRFLDWH3URIHVVRU  $WVXVKL1DNDJDZD' 6FL(LNL<DPDVKLWD3K'

KWWSZZZSURWHLQRVDNDXDFMSUFVISVXSUDFU\VWUHVHDUFKEHDPOLQH
:HDUHZRUNLQJRQGHYHORSPHQWRIQHZPHWKRGRORJLHVIRU;UD\SURWHLQ

FU\VWDOORJUDSK\XVLQJWKHV\QFKURWURQUDGLDWLRQEHDPOLQHIRUELRORJLFDOPDFURPROHFXODU DVVHPEOLHV DW 63ULQJ %/;8  7KLV EHDPOLQH

XWLOL]HV KLJKEULOOLDQW XQGXODWRU UDGLDWLRQ RI 63ULQJ WR FROOHFW KLJK

⡧5,.(1TXDOLW\ GLIIUDFWLRQ GDWD IURP ELRORJLFDO PDFURPROHFXODU DVVHPEO\
FU\VWDOV

5HVHDUFK3URJUDPV'HYHORSPHQWRIKLJKSUHFLVLRQGLIIUDFWLRQGDWDFROOHFWLRQV\VWHPXVLQJ6\QFKURWURQ5DGLDWLRQ

6WUXFWXUDOVWXGLHVRIELRORJLFDOPDFURPROHFXODUDVVHPEOLHV

 3XEOLFDWLRQV






63ULQJ%/;8 EHDPOLQH GHVLJQHG IRU VWUXFWXUH DQDO\VLV RI ODUJHELRORJLFDOPDFURPROHFXODUDVVHPEOLHV+LJDVKLXUD $ <DPDVKLWD ( <RVKLPXUD 0 +DVHJDZD . )XUXNDZD < .XPDVDND 7 8HQR * <DPDPRWR 0
7VXNLKDUD7  $,3&RQI3URF

&U\VWDOVWUXFWXUHVRI2SU1DQG2SU-RXWHUPHPEUDQHIDFWRUVRIPXOWLGUXJWULSDUWLWHHIIOX[SXPSVRI3VHXGRPRQDVDHUXJLQRVD<RQHKDUD5<DPDVKLWD(1DNDJDZD$  3URWHLQV6WUXFWXUH)XQFWLRQDQG%LRLQIRUPDWLFV

$KLJKUHVROXWLRQVWUXFWXUHRISUHPLFUR51$QXFOHDUH[SRUWPDFKLQHU\2NDGD&<DPDVKLWD(/HH6-6KLEDWD
6.DWDKLUD-1DNDJDZD$<RQHGD<7VXNLKDUD7  6FLHQFH

‘0ROHFXODU$QDO\VLV*URXS
3URIHVVRU &RQFXUUHQW 7HFKQLFDO6WDII㻌

7RVKLIXPL7DNDR'6FL-XQLFKL7DNDJL3K' $WVXVKL1DNDJDZD'6FL
1DRNR1RULRND3K'.HLNR.DZDNDPL

㻌7KLVJURXSLVZRUNLQJRQDQDO\VLVRIWKHSULPDU\VWUXFWXUHRISURWHLQV

DQG SHSWLGHV XVLQJ FRQYHQWLRQDO PDVV VSHFWURPHWHUV DQG SHSWLGH
VHTXHQFHUV 7KLV JURXS LV DFFXPXODWLQJ YDULRXV GDWD IRU TXDOLW\㻌FRQWURORISURWHLQVWRPDNHWKHPDSSURSULDWHIRUPROHFXODUDQDO\VLV◦ 48 ◦◦◦ Research Center for State-of-the-Art Functional Protein Analysis

Institute for Protein Research Osaka University‘105*URXS$VVRFLDWH3URIHVVRU㻌<RKHL0L\DQRLUL3K'
\PL\DQRLUL#SURWHLQRVDNDXDFMS 
6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU &RQFXUUHQW   7RVKLKLNR6XJLNL3K'

2XUUHVHDUFKLVFRQFHUQHGZLWKVWUXFWXUDODQGG\QDPLFDODVSHFWVRISURWHLQIXQFWLRQ:H
DUH LQWHUHVWHG LQ HOXFLGDWLQJ FRUUHODWLRQ EHWZHHQ VWUXFWXUH G\QDPLFV DQG ELRORJLFDOIXQFWLRQVRIPDFURPROHFXODUSURWHLQV7RSXUVXHRXUJRDOVZHXVHQXFOHDUPDJQHWLFUHVRQDQFHVSHFWURVFRS\ 105 $WSUHVHQWWKHVROXWLRQ105VWXG\RIODUJHUSURWHLQV !N'D UHOLHVH[FOXVLYHO\XSRQWKHLQIRUPDWLRQREWDLQHGIURPWKHEDFNERQH1+DQGPHWK\O&+VLJQDOVZKLFKDUHQRWVXIILFLHQWIRUSUHFLVHG\QDPLFVDQGVWUXFWXUDODQDO\VLVRI

SURWHLQV ,Q RUGHU WR VROYH WKLV SUREOHP ZH KDYH EHHQ GHYHORSLQJ WKH 6WHUHR$UUD\ ,VRWRSH /DEHOLQJ 6$,/  105PHWKRGV7KHQHZ6$,/105PHWKRGVDOORZXVWRREVHUYHWKHVHYHUDO+&VLJQDOVLQtN'DODUJHPROHFXODUSURWHLQV 7KLV LQIRUPDWLRQ LV YHU\ XVHIXO IRU XQGHUVWDQGLQJ WKH SUHFLVH VWUXFWXUH DQG G\QDPLF SURSHUW\ RI YDULRXVPDFURPROHFXODUSURWHLQVVXFKDVPHPEUDQHSURWHLQVDQGSURWHLQSURWHLQFRPSOH[HV

5HVHDUFK3URJUDPV'HYHORSPHQWRIQHZ105PHWKRGVIRUVWUXFWXUHDQGG\QDPLFVVWXG\RIODUJHUSURWHLQV

4XDQWLWDWLYHDQDO\VLVRIVWUXFWXUDOUHDUUDQJHPHQWIRUSU RWHLQVXSRQLQWHUDFWLQJZLWKRWKHUPROHFXOHV)XUWKHURSWLPL]DWLRQRIVWDEOHLVRWRSHODEHOLQJRI6$,/DPLQRDFLGV3XEOLFDWLRQV
 6WUXFWXUDODQGIXQFWLRQDODQDO\VLVRIWKH&WHUPLQDOUHJLRQRI)OL*DQHVVHQWLDOPRWRUFRPSRQHQWRI9LEULR1D

GULYHQIODJHOOD0L\DQRLUL<+LMLNDWD$1LVKLQR<*RKDUD02QRXH<.RMLPD6.RMLPD&6KLUDL7.DLQRVKR0+RPPD0  6WUXFWXUH

 +LJKO\HIILFLHQWUHVLGXHVHOHFWLYHODEHOLQJZLWKLVRWRSHODEHOHG,OH/HXDQG9DOXVLQJDQHZDX[RWURSKLF (FROLVWUDLQ0L\DQRLUL<,VKLGD<7DNHGD07HUDXFKL7,QRX\H0.DLQRVKR0  -%LRPRO105 8VHRI+'LVRWRSHHIIHFWVWRJDWKHULQIRUPDWLRQDERXWK\GURJHQERQGLQJDQGK\GURJHQH[FKDQJHUDWHV7DNHGD
00L\DQRLUL<7HUDXFKL7<DQJ&-.DLQRVKR0  -0DJQ5HVRQ

 'LIIHUHQWLDO LVRWRSHODEHOLQJ IRU /HX DQG 9DO UHVLGXHV LQ D SURWHLQ E\ ( FROL FHOOXODU H[SUHVVLRQ XVLQJ VWHUHRVSHFLILFDOO\PHWK\OODEHOHGDPLQRDFLGV0L\DQRLUL<7DNHGD02NXPD.2QR$07HUDXFKL7.DLQRVKR0 -%LRPRO105

 0ROHFXODU EDVHV RI PXOWLPRGDO UHJXODWLRQ RI D IXQJDO WUDQVLHQW UHFHSWRU SRWHQWLDO 753  FKDQQHO ,KDUD 0
+DPDPRWR60L\DQRLUL<7DNHGD0.DLQRVKR0<DEH,8R]XPL1<DPDVKLWD$  -%LRO&KHP



 $OWHUQDWLYH 6$,/7US IRU UREXVW DURPDWLFVLJQDO DVVLJQPHQW DQG GHWHUPLQDWLRQ RI WKH Т FRQIRUPDWLRQ E\ LQWUDUHVLGXH12(V0L\DQRLUL<7DNHGD0-HH-2QR$02NXPD.7HUDXFKL7.DLQRVKR0   -%LRPRO105

 0\RVLQ9,XQGHUJRHV FDUJRPHGLDWHGGLPHUL]DWLRQ<X&)HQJ::HL=0L\DQRLUL<:HQ:=KDR<=KDQJ0 &HOO 

◦◦◦ 49 ◦IPR㻌‘(0*URXS$VVRFLDWH3URIHVVRU㻌.HQML,ZDVDNL3K'
LNHQML#SURWHLQRVDNDXDFMS $VVLVWDQW3URIHVVRU    1DR\XNL0L\D]DNL3K'㻌
6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU &RQFXUUHQW   $NLKLUR.DZDPRWR3K'
2XU PLVVLRQ LV WR GHYHORS VLQJOHSDUWLFOH HOHFWURQ FU\RPLFURVFRS\ FU\R(0  DQG SURYLGHKLJKTXDOLW\ GDWD XVLQJ FXWWLQJ HGJH FU\R(0 LQVWUXPHQWV 2XU FU\R(0 IDFLOLW\ KDV WKUHH

HOHFWURQFU\RPLFURVFRSHVDQGRQHFRQYHQWLRQDOWUDQVPLVVLRQHOHFWURQPLFURVFRSH7KHEHVWRIWKHPLV7LWDQ.ULRVHTXLSSHGZLWKUHFHQWDGYDQFHGGHYLFHVZKLFKDUHGLUHFWHOHFWURQFDPHUDV.6XPPLWZLWKDQHQHUJ\ILOWHUDVZHOODV)DOFRQ&VFRUUHFWRU,QDGGLWLRQERWK9ROWDSKDVHSODWH 933 DQGVSKHULFDODEHUUDWLRQFRUUHFWRU &VFRUUHFWRU 

DUHLQVWDOOHGKHUH933LVDSRZHUIXOWRROWRYLVXDOL]HVPDOOELRORJLFDOPROHFXOHV7KLV&VFRUUHFWRULVWKHRQO\RQH

LQVWDOOHGZLWKLQWKHELRORJ\ILHOGLQ-DSDQ7KHDXWRPDWLFLPDJHDFTXLVLWLRQV\VWHPLVDOVRYHU\LPSRUWDQWWRWU\WR
DQDO\]H VWUXFWXUDO YDULDWLRQ RI WDUJHW PROHFXOHV ,Q IDFW RXU 7LWDQ .ULRV DFKLHYHG ̅)6&UHVROXWLRQ RI Ў
JDODFWRVLGDVH  ZKLFKLVWKHKLJKHVWUHVROXWLRQLQ-DSDQDQGDZRUOGOHDGLQJDGYDQFH

7LWDQ .ULRV$XWRPDWLF'DWD$FTXLVLWLRQ㻌

1HDUDWRPLFVWUXFWXUHV㻌 Å㻌

㸨㻌 Å

㸨㻌㻌

5HVHDUFK3URJUDPV'HYHORSPHQWRIVLQJOHSDUWLFOHFU\R(0PHWKRGVIRUGUXJGLVFRYHU\DQGGHYHORSPHQW$QDO\VLVRIVWUXFWXUDOSRO\PRUSKLVPRIWDUJHWPROHFXOHV'HYHORSPHQWRIWKHPHWKRGVIRUVWUXFWXUDOGHWHUPLQDWLRQRID VPDOOSURWHLQXVLQJFU\R(03XEOLFDWLRQVέϧ΢Ψୱཽࢢմੵ๑Ͷ͕͜Ζύʖχ໚Ͳ͹਒ิ ‫ݩࣰ   ࣑ݓ࡜آ‬ҫָ  ೧  ݆ߺ 9RO 1R

SS

1DWXUDODQG$UWLILFLDO3URWHLQ&DJHV'HVLJQ6WUXFWXUHDQG7KHUDSHXWLF$SSOLFDWLRQV+HGGOH-*&KDNUDERUWLόʖϗϩৈΝಣΊմ͚Կָৈʰߑଆմੵ͹ͪΌ͹έϧ΢Ψుࢢ‫ݨ‬ඏ‫ں‬͹֋൅ʱ‫   ࣑ݓ࡜آ‬Կָ ೧  ݆' K\EULG DQDO\VLV $SSURDFK IRU ELQGLQJ WKUHHGLPHQVLRQDO DWRPLF PRGHO E\ HOHFWURQ PLFURVFRS\ LPDJH8QGHUVWDQGLQJWKH$VVHPEO\RIDQ$UWLILFLDO3URWHLQQDQRWXEH1DJDQR6%DQZHOO(),ZDVDNL.0LFKDODN6 DQG,ZDVDNL.  &XUU2SLQ6WUXFW%LRO
ߺ9ROSSPDWFKLQJ0DWVXPRWR$0L\D]DNL17DNDJL-DQG,ZDVDNL.  6FL5HS

03DOND5=KDQJ.<-9RHW$5'DQG+HGGOH-*  $GY0DWHU,QWHUIDFHV

+LUDED\DVKL.<XGD(7DQDND1.DWD\DPD6,ZDVDNL.0DWVXPRWR7.XULVX2XWWHQ*):)XNX\DPD.7DNDKDVKL<DQG:DGD.)XQFWLRQDO'\QDPLFV5HYHDOHGE\WKH6WUXFWXUHRIWKH6XI%&'&RPSOH[D1RYHO$73ELQGLQJ&DVVHWWH $%& 3URWHLQ7KDW6HUYHVDVD6FDIIROGIRU,URQ6XOIXU&OXVWHU%LRJHQHVLV  -%LRO&KHP

7VXNDVDNL<0L\D]DNL10DWVXPRWR$1DJDH6<RQHPXUD67DQRXH7,ZDVDNL.DQG7DNHLFKL0*LDQWFDGKHULQV)DWDQG'DFKVRXVVHOIEHQGWRRUJDQL]HSURSHUO\VSDFHGLQWHUFHOOXODUMXQFWLRQV   3URF

1DWO$FDG6FL86$  

◦ 50 ◦◦◦ Research Center for State-of-the-Art Functional Protein Analysis

Institute for Protein Research Osaka University

㻌/DERUDWRU\ RI 3URWHLQ 'DWDEDVHV
㻌/DERUDWRU\RI3URWHLQ'DWDEDVHV‘3'%M 3URWHLQ'DWD%DQN-DSDQ *URXS
3URIHVVRU &RQFXUUHQW $VVRFLDWH3URIHVVRU

6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU
6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU*HQML.XULVX3K'$NLUD5.LQMR3K' 
7DNHVKL.DZDEDWD3K'+LURIXPL6X]XNL3K'*HUW-DQ%HNNHU3K' &RQFXUUHQW :HWKH3'%M 3URWHLQ'DWD%DQN-DSDQKWWSVSGEMRUJ JURXSFXUDWH
HGLW DQG SURFHVV WKH ' DWRPLF VWUXFWXUH GDWD RI ELRORJLFDO
PDFURPROHFXOHV DQG SURYLGHWKHP IUHHO\DQG SXEOLFO\ RQ RXU RZQ :HEVLWH DV RQH RI WKH ZZ3'% PHPEHUV ZRUOGZLGH 3'%
KWWSVZZSGERUJ  :H DOVR SUHSDUH YDULRXV VHUYLFHV IRU UHVHDUFKHUVDQGVWXGHQWVZKRDUHLQWHUHVWHGLQVWUXFWXUDOELRORJ\PROHFXODUJUDSKLFVYLHZHU0ROPLOPROHFXODUVXUIDFHGDWDEDVHIRUIXQFWLRQDOVLWHVH)VLWH
DQG(01DYLJDWRULPDJHYLHZHUIRUWKHVWUXFWXUHVE\HOHFWURQPLFURVFRS\

3XEOLFDWLRQV.LQMR $5 %HNNHU *- 6X]XNL + 7VXFKL\D < .DZDEDWD 7 ,NHJDZD < 1DNDPXUD + 3URWHLQ 'DWD %DQN -DSDQ
3'%M XSGDWHGXVHULQWHUIDFHVUHVRXUFHGHVFULSWLRQIUDPHZRUNDQDO\VLVWRROVIRUODUJHVWUXFWXUHV  1XFO$FLGV5HV''.LQMR$5<DPDVKLWD51DNDPXUD+3'%M0LQH'HVLJQDQGLPSOHPHQWDWLRQRIUHODWLRQDOGDWDEDVHLQWHUIDFHIRU
3URWHLQ'DWD%DQN-DSDQ  'DWDEDVHEDT

‘3'%M%05% %LR0DJ5HV%DQN *URXS
 3URIHVVRU &RQFXUUHQW 

7RVKLPLFKL)XMLZDUD'6FL
 6SHFLDOO\DSSRLQWHG$VVRFLDWH3URIHVVRU 1DRKLUR.RED\DVKL3K'
:HWKH3'%M%05% KWWSEPUEGHSSGEMRUJ JURXSFROOHFWDQGSURFHVVWKH105H[SHULPHQWDOGDWDRIELRORJLFDOPROHFXOHVDQGSURYLGHWKHPRQRXU:HEVLWHFROODERUDWLQJZLWK%05%DW8QLYHUVLW\RI:LVFRQVLQ0DGLVRQLQ86$%05%FROOHFWVDQQRWDWHVDUFKLYHVDQGGLVVHPLQDWHV ZRUOGZLGHLQWKHSXEOLFGRPDLQ LPSRUWDQWVSHFWUDODQGTXDQWLWDWLYHGDWDGHULYHGIURP105 VSHFWURVFRSLF LQYHVWLJDWLRQV RI ELRORJLFDO PDFURPROHFXOHV DQG
PHWDEROLWHV 7KH JRDO LV WR HPSRZHU VFLHQWLVWV LQ WKHLU DQDO\VLV RI WKHVWUXFWXUH G\QDPLFV DQG FKHPLVWU\ RI ELRORJLFDO V\VWHPV DQG WR VXSSRUW㻌IXUWKHUGHYHORSPHQWRIWKHILHOGRIELRPROHFXODU105VSHFWURVFRS\3XEOLFDWLRQV<RNRFKL0.RED\DVKL18OULFK(/.LQMR$5,ZDWD7,RDQQLGLV<(/LYQ\00DUNOH\-/   1DNDPXUD+.RMLPD&)XMLZDUD73XEOLFDWLRQRIQXFOHDUPDJQHWLFUHVRQDQFHH[SHULPHQWDOGDWDZLWKVHPDQWLFZHEWHFKQRORJ\DQG
WKHDSSOLFDWLRQWKHUHRIWRELRPHGLFDOUHVHDUFKRISURWHLQV  -%LRPHG6HPDQWLFV 
.RED\DVKL1+DUDQR<7RFKLR11DNDWDQL(.LJDZD7<RNR\DPD60DGLQJ68OULFK(/0DUNOH\-/$NXWVX+)XMLZDUD7$QDXWRPDWHGV\VWHPGHVLJQHGIRUODUJHVFDOH105GDWDGHSRVLWLRQDQGDQQRWDWLRQ$SSOLFDWLRQWR RYHU  DVVLJQHG FKHPLFDO VKLIW GDWD HQWULHV WR WKH %LR0DJ5HV%DQN IURP WKH 5,.(1 6WUXFWXUDO*HQRPLFV3URWHRPLFV,QLWLDWLYHLQWHUQDOGDWDEDVH  -%LRPRO105

‘&6'
&DPEULGJH6WUXFWXUDO'DWDEDVH *URXS 3URIHVVRU &RQFXUUHQW   *HQML.XULVX3K'

&6' 7KH&DPEULGJH6WUXFWXUDO'DWDEDVH LVDGDWDEDVHIRUVPDOOPROHFXOHFU\VWDOVWUXFWXUHVFRQVWUXFWHGE\&&'&
7KH&DPEULGJH&U\VWDOORJUDSKLF'DWD&HQWUH :HVXSSRUWDFDGHPLFUHVHDUFKHUVLQ-DSDQDVWKH&&'&bV1DWLRQDO$IILOLDWHG&HQWUH 1$& DQGSURYLGHD&6'SRUWDOWRWKHXVHUVRI2VDND8QLYHUVLW\XQGHUWKHFDPSXVOLFHQVH.

◦◦◦ 51 ◦

IPR
㻌

S
S
\
㻌2SHQ6SDFH/DERUDWRU\IRU$GYDQFHG3URWHLQ6FLHQFH㻌㻌㻌
9LVLWLQJ3URIHVVRU

7DNDVKL6DWRK3K'
WDNDVKLVDWRZ#ULJDNXFRP 6WXGLHVRQPHWKRGLPSURYHPHQWVLQ;UD\FU\VWDOORJUDSK\DQGUHODWHGDUHDOLNHFU\R7(0H[SHULPHQWVWRSXVKWKHOLPLWVDZD\IURPWKHFXUUHQWO\DYDLODEOHOHYHOV7KHVWXG\ZLOOEHDWWKHFORVHO\SUDFWLFDOOHYHOVDVLQV\VWHPLQWHJUDWLRQLQQRYDWLRQVDQGVRIWZDUHLPSOHPHQWDWLRQVWRVKRZWKHDFWXDOHIIHFWLYHQHVVDQG
LPSRUWDQFH

3ULQFLSOHDQGPHWKRGVRI;UD\FU\VWDOORJUDSK\ZHUHHVWDEOLVKHGPRUHWKDQGHFDGHVKRZHYHUWKHUHDUH
VWLOO QXPEHUV RI dLQFRQYHQLHQW WUXWKVb WKDW KDYH QRW EHHQ FRQVLGHUHG DQG GLVFXVVHG ZHOO EHFDXVH WKHDYDLODEOHUHVXOWVZHUHHQRXJKWRJLYHWKHHYLGHQFHDWWKHVFLHQWLILFVWXGLHVDWWKDWGD\V1HYHUWKHOHVVWKH
SDUWRIWKRVHLVVXHVKDVOLPLWHGWKHTXDOLW\RIWKHUHVXOWVDQGWKXVWKHVFLHQWLILFDFKLHYHPHQWVZHUHOLPLWHG
DWDFHUWDLQOHYHODQGWKDWKDYHFRQVWLWXWHGDFRPPRQVHQVHLQWKHILHOG +RZHYHUIRUH[DPSOHWHFKQRORJ\LPSURYHPHQWVDWVPDOOPROHFXODUFU\VWDOORJUDSK\DUHQRZWRHQDEOHWKHOHYHORI 5LQWLQGDLO\PLQXWH VH[SHULPHQWVDQGDWWKHPDQ\FDVHVERQGLQJHOHFWURQGHQVLWLHVDUH
LQWHUIHULQJ WKH FRQYHUJHQFH RI 5 RQFH SHRSOH FROOHFWHG WKH GDWD WR IXOILOO WKH ,8&U UHTXLUHPHQWV :H㻌
IRXQGWKDWJUHDWHQKDQFHPHQWRIWKHGDWDTXDOLWLHVLQWKHSURWHLQFU\VWDOORJUDSK\FRXOGEHDFKLHYHGE\XWLOL]LQJWKHWHFKQRORJLHVLQ60FU\VWDOORJUDSK\)RUH[DPSOHGUDPDWLFLPSURYHPHQWVLQHOHFWURQGHQVLW\IRUSKDUPDFHXWLFDOSURWHLQFU\VWDOORJUDSK\WKDWHQDEOHWRVKRZPXFKUHOLDEOHGUXJGHVLJQWHPSODWHDQG
WRJLYHPXFKHDVLHUSKDVLQJDW66$'H[SHULPHQWVWKDWH[SDQGWKHXWLOLW\RIWKHNQRZQEXWPHUHO\XVHGPHWKRG:HZLOODOVRZRUNRQZHWDVSHFWVRIWKHPHWKRGVDVZHOOWRPDNHWKHLPSURYHPHQWVDSSOLFDEOHLQWRUHDOVFLHQWLILFVWXG\VLWXDWLRQV

5HVHDUFK3URJUDPV

8WLOL]DWLRQRIWKHWHFKQRORJLHVDWWKHODERUDWRU\GLIIUDFWLRQH[SHULPHQWVDQGVWUXFWXUHDQDO\VHVLQWR
V\QFKURWURQUDGLDWLRQH[SHULPHQWV

1RYHOPHWKRGVWRLPSURYHVLQJOHSDUWLFOHDQDO\VLVLQFU\R7(0H[SHULPHQWVXWLOL]LQJWKHLQIRUPDWLRQREWDLQHGLQGLIIUDFWLRQH[SHULPHQWV

0HWKRGGHYHORSPHQWIRUGHQRYRVWUXFWXUHDQDO\VHVXWLOL]LQJFRFU\VWDOOL]DWLRQIDFWRUV3XEOLFDWLRQV$EVROXWH VWUXFWXUH GHWHUPLQDWLRQ RI FRPSRXQGV ZLWK D[LDO DQG SODQDU FKLUDOLW\ XVLQJ WKH FU\VWDOOLQH VSRQJHPHWKRG6KRWD<RVKLRND<DVXKLGH,QRNXPD0DQDEX+RVKLQR7DNDVKL6DWRDQG0DNRWR)XMLWD&KHP6FL
$IILQLW\7UDS 3RO\DFU\ODPLGH *HO (OHFWURSKRUHVLV $ 1RYHO 0HWKRG RI &DSWXULQJ 6SHFLILF 3URWHLQV E\ (OHFWUR
7UDQVIHU&KLKLUR$ZDGD7DNDVKL6DWRDQG7RVKLIXPL7DNDR$QDO&KHP

6WUXFWXUDO IHDWXUHV RI (VFKHULFKLD FROL KHDWVWDEOH HQWHURWR[LQ WKDW DFWLYDWHV PHPEUDQHDVVRFLDWHG JXDQ\O\O
F\FODVH7DNDVKL6DWRDQG<DVXWVXJX6KLPRQLVKL(XU-$OOHUDQG&OLQ,PPXQ

◦ 52 ◦◦◦ Research Center for State-of-the-Art Functional Protein Analysis

Technology Division

IPR
㻌
㻌

7HFKQRORJ\'LYLVLRQ㻌KWWSZZZSURWHLQRVDNDXDFMSJLMXWVXLQGH[KWPO㻌

7HFKQLFDO6WDII㻌.HLNR.DZDNDPL 7DNDJL/DE   ([Wʥ NHLNR#SURWHLQRVDNDXDFMS㻌 㻌7DNDVKL.RVDGD㻌㻌㻌'DWDEDVH/DE([Wʥ WDNDVLˑSURWHLQRVDNDXDFMS 1DRNR1RULRND.XULVX/DE  ([Wʥ QDRNR#SURWHLQRVDNDXDFMS

1DR\XNL$EH

)XMLZDUD/DE ([W  QBDEH#SURWHLQRVDNDXDFMS

7RVKLQRUL7VXMLL)XUXNDZD/DE([Wʥ WRVKLQRULWVXMLL#SURWHLQRVDNDXDFMS

㻌
㻌



$FWLYLW\5HSRUW㻌㻌ʀ$QLPDO)DFLOLW\ 7VXMLLʥ
0DQDJHPHQWDQGPDLQWHQDQFHLQDQDQLPDOIDFLOLW\ 63)PLFHUDWV 㻌7HFKQLFDOJXLGDQFHIRUH[SHULPHQWDODQLPDOKDQGOLQJPHWKRGV㻌

3URGXFWLRQRIJHQHWLFDOO\PRGLILHGPLFH

3URGXFWLRQRIPRXVHIUR]HQHPEU\RVUHVWRUDWLRQRIPRXVHIURPIUR]HQHPEU\RV㻌㻌 ʀ5DGLRLVRWRSH)DFLOLWLHV $EH 㻌㻌    0DQDJHPHQWDQGPDLQWHQDQFHRIWKH5,)DFLOLWLHV2SHUDWLRQDQGPDLQWHQDQFHRIWKHLQVWUXPHQWVLQWKH5,IDFLOLWLHV        0DQDJHPHQWRIUDGLDWLRQZRUNHUV SHRSOHLQ 㻌    㻌 ʀ*HQHUDO3XUSRVH&RPSXWHU6\VWHPDQG+LJKSHUIRUPDQFH;UD\'LIIUDFWRPHWHUVʤ.RVDGDʥ㻌        8VHUVRI&38VHUYHUVLQIURP,35IURPRWKHUXQLYHUVLWLHVIURPDQRWKHULQVWLWXWH㻌㻌 ʀ/DERUDWRU\RI$GYDQFHG3URWHLQ&KDUDFWHUL]DWLRQ  0ROHFXODU$QDO\VLV*URXS1RULRND.DZDNDPLʥ㻌    2SHUDWLRQDQGPDLQWHQDQFHRIWKHLQVWUXPHQWVLQWKHPROHFXODUDQDO\VLVODERUDWRU\ 5RRP    㻌    $QDO\VLVVHUYLFHIRUSURWHLQVHTXHQFLQJ       $%,3URFLVHF/&    ʁ 2VDND8QLYHUVLW\ ,6,5(QJLQHHULQJ                     2WKHUV 7RN\R0HWURSROLWDQ,QVWLWXWHRI0HGLFDO6FLHQFH+RVKL8QLY+LURVKLPD8QLY.DQD]DZD8QLY(KLPH8QLY<RNRKDPD&LW\8QLY.LQGDL8QLY       6KLPDG]X33643364JUDGLHQW   ʁVDPSOHVLQWRWDODW,35

ʀ:RUNDWHDFKODERUDWRU\

1◦ 54 ◦◦◦ Technology Division

Division of DonatedFund Research

Division of Matrixome Research and Application

IPR
㻌
㻌

RI0DWUL[RPH5HVHDUFKDQG$SSOLFDWLRQ
0DWUL[RPH 5HVHDUFK DQG $SSOLFDWLRQ㻌'LYLVLRQ RI㻌㻌㻌㻌
3URIHVVRU
 .L\RWRVKL6HNLJXFKL3K'㻌㻌㻌㻌㻌
VHNLJXFK#SURWHLQRVDNDXDFMS 㻌㻌

$VVLVWDQW3URIHVVRU

.D]XPDVD)XMLWD3K'㻌
2XUORQJWHUPJRDOLVWRXQGHUVWDQGWKHPROHFXODUPHFKDQLVPVGHILQLQJPRUSKRJHQHWLFLQWHUDFWLRQVRIFHOOV ZLWK WKHLU VXUURXQGLQJ PLFURHQYLURQPHQW LH H[WUDFHOOXODU PDWULFHV )LJ   :H DUH SDUWLFXODUO\㻌LQWHUHVWHGLQWKHUROHVRIWKHEDVHPHQWPHPEUDQHLQKLVWRJHQHVLVRUJDQRJHQHVLVZLWKDQHPSKDVLVRQ㻌WKH PROHFXODU LQWHUDFWLRQV RI EDVHPHQW PHPEUDQH SURWHLQV ZLWK WKHLU UHFHSWRUV RQ WKH FHOO VXUIDFH DV
㻌ZHOODVWKHUHVXOWLQJVLJQDOLQJHYHQWVWKDWUHJXODWHSUROLIHUDWLRQGLIIHUHQWLDWLRQDSRSWRVLVDQGPRWLOLW\RI㻌FHOOV(PSKDVLVLVDOVRJLYHQWRWKHIDEULFDWLRQRIFXOWXUHVXEVWUDWHVFXVWRPL]HGIRULQGLYLGXDOFHOOW\SHVLQFOXGLQJSOXULSRWHQWDQGWLVVXHVWHPFHOOVWRSURPRWHVWHPFHOOUHVHDUFKDQGUHJHQHUDWLYHPHGLFLQH

㻌

㻌






)LJ  5HJXODWLRQ RI FHOO JURZWKGLIIHUHQWLDWLRQ DQG VXUYLYDO E\
H[WUDFHOOXODUPDWUL[ 5HVHDUFK3URJUDPV &RPSUHKHQVLYHLPPXQRKLVWRFKHPLFDOSURILOLQJ RIEDVHPHQWPHPEUDQHSURWHLQV 










5HJXODWRU\PHFKDQLVPVRIFHOOXODUIXQFWLRQVE\H[WUDFHOOXODUPDWULFHV
0ROHFXODUEDVLVRIH[WUDFHOOXODUPDWUL[UHFRJQLWLRQE\LQWHJULQV

3K\VLRORJLFDOIXQFWLRQVRISRO\GRPLQYDVFXODUGHYHORSPHQW'HYHORSPHQWRIFXOWXUHVXEVWUDWHVIRUHPEU\RQLFDQGWLVVXHVWHPFHOOV
3XEOLFDWLRQV5HFRPELQDQWODPLQLQIUDJPHQWVHQGRZHGZLWKFROODJHQELQGLQJDFWLYLW\DWRROIRUFRQIHUULQJODPLQLQOLNHFHOO㻌

0HFKDQLVWLFEDVLVIRUWKHUHFRJQLWLRQRIODPLQLQE\DELQWHJULQ7DNL]DZD0$ULPRUL77DQLJXFKL<.LWDJR
㻌

㻌









DGKHVLYHDFWLYLW\WRFROODJHQPDWULFHV6DWR1LVKLXFKL5/L6(ELVX)6HNLJXFKL.  0DWUL[%LRO

<<DPDVKLWD(7DNDJL-6HNLJXFKL.  6FL$GYH

7KUHHGLPHQVLRQDO FHOO VKDSHV DQG DUUDQJHPHQWV LQ KXPDQ VZHDW JODQGV DV UHYHDOHG E\ ZKROHPRXQWLPPXQRVWDLQLQJ.XUDWD5)XWDNL61DNDQR,)XMLWD)7DQHPXUD$0XURWD+.DWD\DPD,2NDGD)6HNLJXFKL.
 3/R62QHH

3RO\GRPLVDQH[WUDFHOOXODUPDWUL[SURWHLQLQYROYHGLQO\PSKDWLFYHVVHOUHPRGHOLQJ0RURRND1)XWDNL66DWR

1LVKLXFKL51LVKLQR07RWDQL<6KLPRQR&1DNDQR,1DNDMLPD+0RFKL]XNL16HNLJXFKL.  &LUF5HV

0ROHFXODUEDVLV RI WKHOLJDQG ELQGLQJ VSHFLILFLW\RI DYE LQWHJULQ 2]DZD $ 6DWR < ,PDED\DVKL 7 8HPXUD 7

7DNDJL-6HNLJXFKL.  -%LRO&KHP

0ROHFXODUEDVLVRIODPLQLQLQWHJULQLQWHUDFWLRQV<DPDGD0DQG6HNLJXFKL.   &XUU7RS0HPEU 


%DVHPHQWPHPEUDQHDVVHPEO\RIWKHLQWHJULQDEOLJDQGQHSKURQHFWLQUHTXLUHV)UDVHUV\QGURPHDVVRFLDWHG

SURWHLQV.L\R]XPL'7DNHLFKL01DNDQR,6DWR<)XNXGD76HNLJXFKL.  -&HOO%LRO

㻌
◦ 56 ◦◦◦ Division of Donated Fund ResearchAbout IPR

Educational Activities
Research Activities
Joint-Usage / Research Center
IPR Saff
Administrative Council
Staff and Accounting Data
Floor PlanIPR
(GXFDWLRQDO$FWLYLWLHV$FDGHPLF VWDII RI ,35 HQJDJHV LQ JUDGXDWH FRXUVH HGXFDWLRQ LQ FRRSHUDWLRQ ZLWK WKH *UDGXDWH 6FKRROV RI 6FLHQFH
0HGLFLQHDQG)URQWLHU%LRVFLHQFHV,QRUGHUIRUVWXGHQWVWRVWXG\DQGMRLQWKHUHVHDUFKDW,35WKH\DUHUHTXLUHGWRWDNHFRXUVHVDWWKHLUHQUROOHGGHSDUWPHQWSURYLGHGE\,35DFDGHPLFVWDII$SSUR[LPDWHO\VWXGHQWVLQFOXGLQJLQWHUQDWLRQDOVWXGHQWVDUHVWXG\LQJDW,35HYHU\\HDU

1XPEHURIVWXGHQWVDW,35






*UDGXDWHVFKRRORI)URQWLHU%LRVFLHQFHV

6FKRRO*UDGXDWH
6FKRRORI0HGLFLQH
6FKRRO*UDGXDWH
6FKRRORI6FLHQFH

















8QGHUJUDGXDWH 0DVWHU&RXUVH'RFWRU&RXUVH*UDGXDWHFRXUVHVE\,35' HS DUW PHQ W RI%LRORJLFDO6FLHQ FHV/DERI3URWHLQ2UJDQLF&KHPLVWU\/DERI&HOO6\VWHPV/DERI1DQRELRORJ\/DERI3URWHLQ)ROGLQJ
/DERI0ROHFXODU%LRSK\VLFV/DERI3URWHLQ&U\VWDOORJUDSK\/DERI0HPEUDQH3URWHLQ&KHPLVWU\/DEIRU0ROHFXODUDQG'HYHORSPHQWDO%LRORJ\/DERI*HQRPHDQG&KURPRVRPH)XQFWLRQV
/DERI1XFOHDU1HWZRUN
/DERI+RPHRVWDWLF,QWHJUDWLRQ*UDG X DW H6FK RRO/DERI3URWHLQ3URILOLQJDQG)XQFWLRQDO3URWHRPLFV

RI6FLHQ FH/DERI3URWHLQ6\QWKHVLVDQG([SUHVVLRQ/DERI6XSUDPROHFXODU&U\VWDOORJUDSK\/DEIRU$GYDQFHG%UDLQ)XQFWLRQV/DERI2UJDQHOOH%LRORJ\' HS DUW PHQ W RI&K HPLV W U\
/DERI3URWHLQ2UJDQLF&KHPLVWU\/DERI0ROHFXODU%LRSK\VLFV/DERI3URWHLQ3URILOLQJDQG)XQFWLRQDO3URWHRPLFV' HS DUW PHQ W RI0DFURPROHFX ODU6FLHQ FH/DERI3URWHLQ)ROGLQJ
/DERI3URWHLQ&U\VWDOORJUDSK\/DERI6XSUDPROHFXODU&U\VWDOORJUDSK\*UDG X DW H6FK RRO
RI0HG LFLQ H*UDG X DW H6FK RRORI)URQ W LHU%LRV FLHQ FHV◦ 58 ◦◦◦ About IPR/DEIRU0ROHFXODUDQG'HYHORSPHQWDO%LRORJ\/DEIRU$GYDQFHG%UDLQ)XQFWLRQV/DERI6XSUDPROHFXODU&U\VWDOORJUDSK\/DERI$GYDQFHG3URWHLQ&KDUDFWHUL]DWLRQ 105*URXS/DERI3URWHLQ3URILOLQJDQG)XQFWLRQDO3URWHRPLFV/DERI3URWHLQ6\QWKHVLVDQG([SUHVVLRQ/DEIRU0ROHFXODUDQG'HYHORSPHQWDO%LRORJ\*UDGXDWH3URJUDPIRU
$GYDQFHG,QWHUGLVFLSOLQDU\6WXGLHV
RI2VDND8QLYHUVLW\$ 3URJUDP IRU 6WDWHRIWKH$UW 3URWHLQ 6WUXFWXUDO$QDO\VLV E\ ,35 VWDUWHG LQ  DV RQH RI WKH*UDGXDWH 3URJUDPV IRU $GYDQFHG ,QWHUGLVFLSOLQDU\6WXGLHV RI 2VDND 8QLYHUVLW\ $FDGHPLF VWDII DW ,35
DQG LQYLWHG OHFWXUHUV IURP LQGXVWU\ DQG RWKHULQVWLWXWHV KDYH SURYLGHG JUDGXDWH VWXGHQWV ZLWKOHFWXUHV RQ WKH FXWWLQJHGJH VWUXFWXUDO ELRORJ\ DV
ZHOO DV VRSKLVWLFDWHG ODUJHVFDOH HTXLSPHQW DQGLQWHUQDWLRQDOGDWDEDVHVWKDWKDYHEHHQGHYHORSHGDW
,35

Institute for Protein Research Osaka University

5HVHDUFK$FWLYLWLHV
,35ZDVIRXQGHGE\PHPEHUVLQ)DFXOW\RI6FLHQFHDQG0HGLFDOVFKRRORI2VDND8QLYHUVLW\FRYHULQJGLIIHUHQWILHOGV$FKLHYHPHQWVRI VFLHQFHV VXFK DV FKHPLVWU\ SK\VLFV ELRORJ\ DQG PHGLFLQH 6LQFH WKHQ SURWHLQ UHVHDUFK LQ ,35 KDV PDGH D

UHPDUNDEOHSURJUHVVE\HOXFLGDWLQJVWUXFWXUHVDQGIXQFWLRQVRISURWHLQVDQGE\XQGHUVWDQGLQJWKHLUELRORJLFDOUROHV
IURP WKH PROHFXODU OHYHO WR WKH FHOOXODU DQG WKH KLJKHU OHYHOV )XUWKHUPRUH ,35 KDV EHHQ JRLQJ IRUZDUG WR WKH
LQWHJUDWLYHVWUXFWXUDO OLIH VFLHQFHDV WKH EDVLFVFLHQFH E\ LQYLWLQJ VSHFLDOLVWVLQ WKH ILHOGV RI V\VWHPV ELRORJ\DQG
VLQJOHPROHFXOHDQDO\VLV

5HFHQWDFKLHYHPHQWV







0DMRUUHVHDUFKSURMHFWVJUDQWHGE\JRYHUQPHQWRUJDQL]DWLRQV
ˠ/LVWHGDUHSURMHFWVLQSURJUHVVDVRI)<

ZLWKJUDQWVRIWHQPLOOLRQ\HQDQGRYHU*UDQWVLQ$LGIRU6FLHQWLILF5HVHDUFK .$.(1+,2UJDQL]DWLRQ

3URJUDP)XQGIRUWKH3URPRWLRQRI-RLQW,QWHUQDWLRQDO5HVHDUFK )RVWHULQJ-RLQW,QWHUQDWLRQDO5HVHDUFK*UDQWLQ$LGIRU6FLHQWLILF5HVHDUFK$-636

3URMHFW%RWWRPXSDQGWRSGRZQDSSURDFKHVLQWKHKLHUDUFK\RIELRORJLFDO
V\VWHPVWRVWXG\VLQJOHFHOOWKHUPRG\QDPLFV6WUXFWXUDOHOXFLGDWLRQRIDULVNIDFWRUIRU$O]KHLPHU V'LVHDVH
VRU/$H[WUDFHOOXODUUHJLRQʷ
ʷ

6WUXFWXUDOPHFKDQLVPRIWKH:QWUHFHSWRULQWHUDFWLRQʷ

0HWDEROLFDGDSWDWLRQRILQIODPPDWRU\GLVHDVHV

ʷ

0HFKDQLVPDQGUHJXODWLRQRISURWHLQDJJUHJDWLRQLQFRPSOLFDWHG*UDQWLQ$LGIRU6FLHQWLILF5HVHDUFK FHOOXODUV\VWHPVRQ,QQRYDWLYH$UHDV 5HVHDUFKLQD 6WUDWHJLFGHVLJQRIWKHSURWRQPRWLYHIRUFHEDVHGRQWKHVWUXFWXUDOSURSRVHGUHVHDUFKDUHD

7HUP

DQDO\VLV
1HXURQDOFLUFXLWPHFKDQLVPVRIUHZDUGJRDOGLUHFWHGEHKDYLRU&RQWURORIFKURPRVRPHIXQFWLRQVE\D[LVORRSVWUXFWXUH
FKURPRVRPH' GXULQJPHLRVLV

ʷ
ʷ
ʷ
ʷ

◦◦◦ 59 ◦IPR
2WKHUJRYHUQPHQWDOUHVHDUFKJUDQW
2UJDQL]DWLRQ

3URJUDP

3URMHFW&RUUHODWLYHVWUXFWXUDODQDO\VLVSODWIRUPIRUGUXJGLVFRYHU\DQGOLIH
VFLHQFHV$0('

3ODWIRUP3URMHFWIRU6XSSRUWLQJ'UXJ 5HFRPELQDQWSURGXFWLRQRIKLJKYDOXHWDUJHWSURWHLQVDQGWKHLU'LVFRYHU\DQG/LIH6FLHQFH5HVHDUFK ELQGHUVYLDVWUXFWXUHEDVHGSURWHLQGHVLJQ'DWDVFLHQFHDQGFROODERUDWLYHVXSSRUWDFFHOHUDWLQJLQQRYDWLYH
GUXJGLVFRYHU\DQGOLIHVFLHQFHUHVHDUFK

7HUP
ʷ
ʷ
ʷ(QWUXVWHGUHVHDUFKSURMHFWV2UJDQL]DWLRQ0(;73URJUDP

3URMHFW1053ODWIRUP

0(;7ˢ5,.(13URJUDPIRUFRRUGLQDWLRQWRZDUG
LQWHJUDWLRQRIUHODWHGGDWDEDVHVRUJDQLVP'HYHORSPHQWRILQQRYDWLYHPDJQHWLFLPDJHVHQVRUVEDVHGRQ
FDUERQQDQRHOHFWURQLFV-67(OXFLGDWLRQRIG\QDPLFDOVWUXFWXUHVRIPHPEUDQHYROWDJHVHQVRU&5(676WUXFWXUDOVWXGLHVRIWKHSURWHLQVZRUNLQJLQQRYHOFHOOPHPEUDQH
SRWHQWLDOVLJQDO
6WUXFWXUDOVWXGLHVRQWKHWUDQVLHQWPDFURPROHFXODUFRPSOH[HVIRUPHGXSRQSKRWRDFFOLPDWLRQ'HYHORSPHQWRIXOWUDVRQLFDWLRQDVVLVWHGQRQLQYDVLYHPDFKLQHIRU'HYHORSPHQWRIDGYDQFHGPHDVXUHPHQW
DQGDQDO\VLVV\VWHPVGLDJQRVLVRIQHXURGHJHQHUDWLYHGLVHDVHV'HYHORSPHQWRIXOWUDKLJKVHQVLWLW\VSLQFRUUHODWHGKLJKUHVROXWLRQ105VSHFWURPHWHU$0('

3UDFWLFDO5HVHDUFKIRU,QQRYDWLYH&DQFHU 7RWDOSDFNDJHGHYHORSPHQWRIDSSURSULDWHLQQHUFHOO&RQWUROHQYLURQPHQWDODQGYHFWRULPSURYHPHQWIRUWKHHIILFLHQWJHQRPH&\FOLF,QQRYDWLRQIRU&OLQLFDO(PSRZHUPHQW &L&/(















◦ 60 ◦◦◦ About IPR(QKDQFHPHQWRIIXQFWLRQDOLW\DQGLQWHJUDWLYHPDQDJHPHQWRI3'% 
6LQJOHFHOOHQHUJ\SHUWXUEDWLRQDQGTXDQWLILFDWLRQLQDOLYLQJ

35(672

7HUP(TXLSPHQWGHYHORSPHQWIRUKLJKWKURXJKSXWGHWHUPLQDWLRQRI
SURWHLQVWUXFWXUHV









-636-DSDQ6RFLHW\IRUWKH3URPRWLRQRI6FLHQFH
0(;70LQLVWU\RI(GXFDWLRQ&XOWXUH6SRUWV6FLHQFHDQG7HFKQRORJ\-DSDQ-67-DSDQ6FLHQFHDQG7HFKQRORJ\$JHQF\$0('-DSDQ$JHQF\IRU0HGLFDO5HVHDUFKDQG'HYHORSPHQWInstitute for Protein Research Osaka University







-RLQWUHVHDUFKSURMHFWVZLWKSULYDWHSXEOLFVHFWRUV
本館

྽




動物室

8階



疋田研究室



原田研究室
6階



後藤研究室




5階

52+723KDUPDFHXWLFDO&R/WG



藤原研究室
4階

PDBj

'DLLFKL6DQN\R&RPSDQ\$MLQRPRWR&ROQF
岡田研究室





.DR&RUSRUDWLRQ



オルガネラバイオロジー
7KH1LSSRQ6\QWKHWLF&KHPLFDO,QGXVWU\&R/WG
分析実験棟



研究グループ

0DQGRP&RUSRUDWLRQ



2階

2階


先端計測研究室

セミナー室

固体NMR

1階

北條研究室
Foreign PI Lab

 1RWDEOHUHVHDUFKDFWLYLWLHVNMRグループ

高木研究室
1H[WJHQHUDWLRQQDWXUDOSURGXFWVFKHPLVWU\

栗栖研究室



NMR実験棟

7HFKQRORJ\5HVHDUFK$VVRFLDWLRQIRU

1階

溶液NMR

3階

2階

Ｘ線装置室








共通機器室

先端計測研究室

3階

中川研究室


0$75,;20(,QF
篠原研究室

セミナー室



4階

汎用計算機室

7262+$QDO\VLVDQG5HVHDUFK&HQWHU&R/WG








7階






R I 実験室 6KLRQRJL &R/WG
古川研究室







3KDUPDFHXWLFDOFRQVRUWLXPIRUVWUXFWXUHDQDO\VLV

中村研究室









9階




7HUP

蛋白質解析先端研究センター棟-(2//WG

動物室




&RPSDQ\,QVWLWXWLRQ

事務部

大会議室

所長室 中会議室

図書室

共同研究拠点棟
２階



多階層構造
,Q-XO\WKH3'%M 3URWHLQ'DWD%DQN-DSDQKWWSVSGEMRUJ

生命科学研究室セミナー室
高尾研究室
 DWWKH,QVWLWXWHIRU3URWHLQ5HVHDUFK,35 2VDND8QLYHUVLW\VWDUWHG
体内環境統合談話室
講堂
 DFFHSWLQJQHZVWUXFWXUHVIURPUHVHDUFKHUVPDLQO\LQ$VLDQUHJLRQV
1階

蛋白質研究グループ

 7KH3'%MKDVEHFRPHWKHOHDGLQJDUFKLYHRIWKLVUHJLRQDVRQHRI WKHPHPEHUVRIWKHZZ3'% ZRUOGZLGH3'%KWWSVZZSGERUJ -DSDQ
 7KH3'%MKDVEHHQVXSSRUWHGE\2VDND8QLYHUVLW\DQG-67
建物配置
１階
6FLHQFH
DQG
7HFKQRORJ\$JHQF\
,Q
$SULO

1%'&
1DWLRQDO
分析実験棟%LRVFLHQFH 'DWDEDVH共同研究&HQWHU KWWSELRVFLHQFHGEFMS ZDV IRXQGHG
拠点棟
LQ -67 VR DV WR LQWHJUDWH DOPRVW DOO WKH OLIH VFLHQFH GDWDEDVHV LQ
-DSDQ ,Q NMR
DGGLWLRQ 1%'& LWVHOI ZRUNV DV WKH IXQGLQJ DJHQF\ WR実験棟
VXSSRUWJURXSVZKRGHYHORSDQGPDLQWDLQWKHLUGDWDEDVHVLQYDULRXV
本 館
ILHOGVRIOLIHVFLHQFH3'%MKDVVXFFHHGHGLQEHLQJJUDQWHGZLWKWKH
WLWOH*OREDO&RQVWUXFWLRQDQG,QWHJUDWLRQRI3'%E\1%'&IRUWKH
構造解析,QWHJUDWLRQ3URJUDP
研究棟

月原研究室
株式会社マトリクソーム

加納研究室，三間研究室
マトリクソーム科学（ニッピ）寄附研究部門
マルチスケール構造生物学（日本電子）寄付研究部門

3'%M:HEVLWH:RUOGZLGH3'%$GYLVRU\&RPPLWWHH0HHWLQJ ◦◦◦ 61 ◦IPR -RLQW8VDJHDQG5HVHDUFK&HQWHUIRU3URWHLQV$V D -RLQW 8VDJH DQG 5HVHDUFK &HQWHU RXU PDMRU DFWLYLWLHV DUH WR SURPRWH   MRLQW XVDJH RI DGYDQFHG IDFLOLWLHV DQGHTXLSPHQW DFFHVVWRGDWDEDVHVDQG FROODERUDWLRQVLQFOXGLQJWKRVHOHDGLQJWRWKHGHYHORSPHQWRIKXPDQUHVRXUFHV7KURXJKWKHVHDFWLYLWLHVZHDLPWRDFWLYDWHSURWHLQDQGOLIHVFLHQFHFRPPXQLWLHV
,35PDNHVDQDQQXDOFDOOIRUDSSOLFDWLRQVWRWKHIROORZLQJVHYHQFROODERUDWLYHSURMHFWV  'RPHVWLF&ROODERUDWRUV ,QWHUQDWLRQDO&ROODERUDWLYH5HVHDUFK 6SULQJ&ROODERUDWLYH5HVHDUFK 105&ROODERUDWLYH5HVHDUFK &U\R(0&ROODERUDWLYH5HVHDUFK ,356HPLQDUV ,35)HOORZV

















:HEVLWHRIWKH-RLQW8VDJH5HVHDUFK&HQWHUKWWSZZZSURWHLQRVDNDXDFMSN\RWHQHQ

3RVWHUIRUWKHDQQXDOFDOOIRUDSSOLFDWLRQVˠ5HJXODUDSSOLFDWLRQVDUHUHFHLYHGRQFHSHU\HDUDQG
H[FHSWIRU,35)HOORZVXUJHQWDSSOLFDWLRQVFDQEHDFFHSWHGDOO\HDUDURXQG

5HVHDUFKHUVRQ-RLQW5HVHDUFK3URJUDP
7KH-RLQW5HVHDUFK3URJUDPKDVEHHQHVWDEOLVKHGWRSURYLGHYLVLWLQJVFLHQWLVWVIURPRXWVLGHWKH,35ZKRDUHHQJDJHGLQVWXGLHVRQSURWHLQVZLWKDQRSSRUWXQLW\WRSHUIRUPFRRUGLQDWHGUHVHDUFKDWWKH,35IRUXSWRPRQWKV0RUHWKDQVFLHQWLVWVDUHVHOHFWHG\HDUO\IURPDSSOLFDWLRQVIURPYDULRXVGRPHVWLFLQVWLWXWLRQV
7KH SURJUDP FRYHUVUHVHDUFK DQG WUDYHO H[SHQVHV DQG WKH LQVWLWXWH SURYLGHV IDFLOLWLHV IRU DFFRPPRGDWLRQ 6LQFH WKHEHJLQQLQJ RI SURJUDP LQ  D WRWDO RI  UHVHDUFKHUV KDYH EHHQ DGPLWWHG 5HVHDUFKHUV ZKR ZDQW WR XVH ODUJHLQVWUXPHQWVVXFKDV;UD\DQDO\VLVIDFLOLWLHVVXSHUFRQGXFWLQJPDJQHW105DQG&U\R(OHFWURQ0LFURVFRS\ &U\R(0 
DUHUHTXLUHGWRDSSO\IRUWKLVSURJUDP








◦ 62 ◦◦◦ About IPRInstitute for Protein Research Osaka University,QWHUQDWLRQDO&ROODERUDWLYH 5HVHDUFK 3URJUDP,35HVWDEOLVKHGWKH,QWHUQDWLRQDO&ROODERUDWLYH5HVHDUFK3URJUDPLQLQYLWLQJUHVHDUFKHUVEURDGO\IURPRYHUVHDV8QGHUWKLVSURJUDPUHVHDUFKVKRXOGEHFRQGXFWHGLQWKHIRUPRIDFROODERUDWLRQLQFOXGLQJDWOHDVWRQHRIWKHSULQFLSDOLQYHVWLJDWRUVDW,35RULWVKRXOGXVHSDUWLFXODUH[SHULPHQWDOIDFLOLWLHVRI,35 ,Q)<RYHUVHDVUHVHDUFKHUVIURPFRXQWULHVYLVLWHG,35DQGFRQGXFWHGFROODERUDWLYHUHVHDUFKHVZLWKWKH
SULQFLSDOLQYHVWLJDWRUVDW,35

)DFLOLWLHV

⡧5,.(1%HDPOLQHIRU%LRORJLFDO0DFURPROHFXODU$VVHPEOLHVDW63ULQJ&U\RHOHFWURQ0LFURVFRSHV6ROXWLRQ1056SHFWURPHWHU 
LQDQ8OWUDKLJK0DJQHWLF)LHOG$QDO\WLFDO$SSDUDWXVIRU6XSUDELRPROHFXOHV6ROLGVWDWH1056SHFWURPHWHUV+LJKSHUIRUPDQFH;UD\'LIIUDFWRPHWHU

◦◦◦ 63 ◦IPR,356WDII 'LUHFWRU$VRI0D\

3URIHVVRU$WVXVKL1$.$*$:$'6FL

3URIHVVRU<XML*272'6FL

9LFH'LUHFWRUV

3URIHVVRU

7DNDKLVD)858.$:$0'3K'

3URIHVVRU*HQML.85,683K''LUHFWRURIWKH5HVHDUFK&HQWHU'LYLVLRQRI3URWHLQ&KHPLVWU\'LYLVLRQRI,QWHJUDWHG3URWHLQ)XQFWLRQV/DERUDWRU\RI3URWHLQ2UJDQLF&KHPLVWU\/DERUDWRU\RI*HQRPHDQG&KURPRVRPH)XQFWLRQV

3URIHVVRU+LURQREX+2-23K'

3URIHVVRU$NLUD6+,12+$5$'6FL

$VVRFLDWH3URIHVVRU

7RUX.$:$.$0,3K'$VVRFLDWH3URIHVVRU$VDNR)858.2+5,3K'

$VVLVWDQW3URIHVVRU<X\D$6$+,1$3K'

$VVLVWDQW3URIHVVRU.HQLFKLUR0$768=$.,3K'

$VVLVWDQW3URIHVVRU

0LND+,*$6+,'(3K'/DERUDWRU\RI&HOO6\VWHPV/DERUDWRU\IRU0ROHFXODUDQG'HYHORSPHQWDO%LRORJ\

3URIHVVRU

0DULNR2.$'$3K'

$VVLVWDQW3URIHVVRU.D]XQDUL,:$02723K'

3URIHVVRU

7DNDKLVD)858.$:$0'3K'

$VVLVWDQW3URIHVVRU

6KLJH\XNL0$*,3K'$VVRFLDWH3URIHVVRU<RVKLKLUR2025,0'3K'

$VVLVWDQW3URIHVVRU

6X[LDQJ=+$1*0'3K'$VVLVWDQW3URIHVVRU

7DUR&+$<$0'3K'

$VVLVWDQW3URIHVVRU<XNR68*,7$0'3K'/DERUDWRU\RI1DQRELRORJ\/DERUDWRU\IRU$GYDQFHG%UDLQ)XQFWLRQV

3URIHVVRU<RVKLH+$5$'$3K'$VVRFLDWH3URIHVVRU

0DGRND68=8.,3K'

3URIHVVRU$VVLVWDQW3URIHVVRU+LVDVKL7$'$.80$3K'

$VVLVWDQW3URIHVVRU

7DNDVKL<$0$*8&+,3K'

6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU

7RP0$&3+(56213K'ɻ

7DNDWRVKL+,.,'$0'3K'/DERUDWRU\RI2UJDQHOOH%LRORJ\,QGHSHQGHQW$VVRFLDWH3URIHVVRU

0DVDWR1$.$,'6FL

'LYLVLRQRI3URWHLQ6WUXFWXUDO%LRORJ\/DERUDWRU\IRU1XFOHDU'\QDPLFV
9LVLWLQJ3URIHVVRU

7DNDVKL1$*$120'3K'

3URIHVVRU

7DNDKLVD)858.$:$0'3K'/DERUDWRU\RI3URWHLQ)ROGLQJ
3URIHVVRU<XML*272'6FL

/HFWXUHU<RXQJ+R/(('6FL

6SHFLDOO\DSSRLQWHG/HFWXUHU.HQML6$6$+$5$'6FL

$VVLVWDQW3URIHVVRU

0DVDWRPR62'6FL

6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU.HLLFKL<$0$*8&+,'6FL

/DERUDWRU\RI+RPHRVWDWLF,QWHJUDWLRQ$VVRFLDWH3URIHVVRU

1REXDNL2.8085$3K'

'LYLVLRQRI0XOWLVFDOH,QWHJUDWLYH3URWHLQ6FLHQFH/DERUDWRU\RI1XFOHDU1HWZRUN,QGHSHQGHQW$VVRFLDWH3URIHVVRU-XQNR.$12+3K'/DERUDWRU\RI0ROHFXODU%LRSK\VLFV/DERUDWRU\RI0XOWLVFDOH6WUXFWXUDO%LRORJ\

3URIHVVRU

7RVKLPLFKL)8-,:$5$'6FL

9LVLWLQJ3URIHVVRU&KRMLUR.2-,0$3K'

3URIHVVRU-XQLFKL7$.$*,3K'$VVRFLDWH3URIHVVRU<RK0$768.,3K'

3URIHVVRU*HQML.85,683K'

6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU$\DNR(*$:$3K'$VVRFLDWH3URIHVVRU.HQML,:$6$.,3K'

6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU

7RVKLKLNR68*,.,3K'9LVLWLQJ$VVRFLDWH3URIHVVRU'DPLHQ5+$//3K'

6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU.HQLFKL+$5$'$3K'

$VVLVWDQW3URIHVVRU

7DNDR$5,025,3K'

6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU-XQ)8.$=$:$3K'

$VVLVWDQW3URIHVVRU

1DR\XNL0,<$=$.,3K'

6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU.\RNR)858.,7$3K'

6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU*HUW-DQ%(..(53K'/DERUDWRU\RI3URWHLQ&U\VWDOORJUDSK\/DERUDWRU\IRU,QIHFWLRQ6\VWHPV

3URIHVVRU*HQML.85,683K'

6SHFLDOO\DSSRLQWHG3URIHVVRU<XPLNR,0$,0'3K'$VVRFLDWH3URIHVVRU+LGHDNL7$1$.$3K'

3URIHVVRU

0DULNR2.$'$3K'

6SHFLDOO\DSSRLQWHG$VVRFLDWH3URIHVVRU&KULVWRSK*$5/(3K'

$VVLVWDQW3URIHVVRU

7HWVXNR1$.$1,:$3K'/DERUDWRU\RI0HPEUDQH3URWHLQ&KHPLVWU\,QGHSHQGHQW$VVRFLDWH3URIHVVRU◦ 64 ◦◦◦ About IPR-RML0,0$3K'/DERUDWRU\RI9LVLWLQJ6FLHQWLVWV
9LVLWLQJ3URIHVVRU

7RVKLR<$0$=$.,'6FL

9LVLWLQJ3URIHVVRU,VDR68(7$.(3K'

9LVLWLQJ$VVRFLDWH3URIHVVRU.HQML68*$6(3K'

Institute for Protein Research Osaka University

5HVHDUFK&HQWHUIRU6WDWHRIWKHDUW)XQFWLRQDO3URWHLQ$QDO\VLV/DERUDWRU\RI3URWHLQ3URILOLQJDQG)XQFWLRQDO3URWHRPLFV3URIHVVRU

7RVKLIXPL7$.$2'6FL

'LYLVLRQRI'RQDWHG)XQG5HVHDUFK'LYLVLRQRI0DWUL[RPH5HVHDUFKDQG$SSOLFDWLRQ
3URIHVVRURI'RQDWHG)XQG5HVHDUFK.L\RWRVKL6(.,*8&+,3K'

$VVLVWDQW3URIHVVRURI'RQDWHG)XQG5HVHDUFK/DERUDWRU\RI3URWHLQ6\QWKHVLVDQG([SUHVVLRQ.D]XPDVD)8-,7$3K'

3URIHVVRU-XQLFKL7$.$*,3K'$VVRFLDWH3URIHVVRU.HQML,:$6$.,3K'

$VVLVWDQW3URIHVVRU<X.,7$*23K'

7HFKQLFDO6WDII.HLNR.$:$.$0,$VVLVWDQW3URIHVVRU

7DNDR$5,025,3K'

7HFKQLFDO6WDII

7DNDVKL.26$'$06FL

$VVLVWDQW3URIHVVRU

1DR\XNL0,<$=$.,3K'

7HFKQLFDO6WDII

1DRNR125,2.$3K'

6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU$NLKLUR.$:$02723K'7HFKQLFDO6WDII

1DR\XNL$%(06FL

7HFKQLFDO6WDII

7RVKLQRUL768-,,/DERUDWRU\RI6XSUDPROHFXODU&U\VWDOORJUDSK\

7HFKQRORJ\'LYLVLRQ$GPLQLVWUDWLRQ2IILFH

3URIHVVRU$WVXVKL1$.$*$:$'6FL

$VVRFLDWH3URIHVVRU

0DPRUX68=8.,3K'+HDG$VVRFLDWH3URIHVVRU(LNL<$0$6+,7$3K'*HQHUDO$IIDLUV6HFWLRQ/DERUDWRU\RI$GYDQFHG3URWHLQ&KDUDFWHUL]DWLRQ.RXML2*85$&KLHI

7RPRNR,&+,.,$GPLQLVWUDWLYH6WDII

7DNDKLUR768-,$GPLQLVWUDWLYH6WDII

1RULNR<26+,085$

3URIHVVRU$WVXVKL1$.$*$:$'6FL

3URIHVVRU-XQLFKL7$.$*,3K'

3URIHVVRU

7RVKLIXPL7$.$2'6FL

&KLHI

0LNL85,8$VVRFLDWH3URIHVVRU<RKHL0,<$12,5,3K'

'HSXW\&KLHI,VDR7$.(0272$VVRFLDWH3URIHVVRU(LNL<$0$6+,7$3K'$GPLQLVWUDWLYH6WDII.HQVXNH.,0272$VVRFLDWH3URIHVVRU.HQML,:$6$.,3K'$GPLQLVWUDWLYH6WDII$\XPL.2%$<$6+,

6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU

7RVKLKLNR68*,.,3K'

6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU$NLKLUR.$:$02723K'$FFRXQWLQJ6HFWLRQ

3URMHFW7HDPRI-RLQW8VDJH5HVHDUFK&HQWHU&RPPLVVLRQHG6WDII

0DVDNR6$72

5HVHDUFK6XSSRUW6HFWLRQ/DERUDWRU\RI3URWHLQ'DWDEDVHV&KLHI+LURKDUX0,&+,.,

3URIHVVRU*HQML.85,683K'

'HSXW\&KLHI&KLDNL220$768

3URIHVVRU

7RVKLPLFKL)8-,:$5$'6FL

$GPLQLVWUDWLYH6WDII.D]XVKL7$.$+$0$$VVRFLDWH3URIHVVRU$NLUD.,1-23K'

6SFHLDOO\DSSRLQWHG$GPLQLVWUDWLYH6WDII

6SHFLDOO\DSSRLQWHG$VVRFLDWH3URIHVVRU

1DRKLUR.2%$<$6+,3K'

6SHFLDOO\DSSRLQWHG$VVRFLDWH3URIHVVRU

7DNHVKL.$:$%$7$3K'

6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU+LURIXPL68=8.,3K'

6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU*HUW-DQ%(..(53K'

3XEOLF5HODWLRQV2IILFH<XNR7$1$.$

6SFHLDOO\DSSRLQWHG$GPLQLVWUDWLYH6WDII

7RPRNR6+,0,=8

2SHQ6SDFH/DERUDWRU\IRU$GYDQFHG3URWHLQ6FLHQFH
9LVLWLQJ3URIHVVRU

7DNDVKL6$723K'$GPLQLVWUDWLYH &RXQFLO&KDLUʁ$WVXVKL1DNDJDZD     'LUHFWRU,QVWLWXWHIRU3URWHLQ5HVHDUFK2VDND8QLYHUVLW\2VDPX1XUHNL

3URIHVVRU*UDGXDWH6FKRRORI6FLHQFHWKH8QLYHUVLW\RI7RN\R.HLLFKL1DND\DPD

3URIHVVRU0HGLFDO,QVWLWXWHRI%LRUHJXODWLRQ.\XVKX8QLYHUVLW\+LGH\D)XNX]DZD

3URIHVVRU*UDGXDWH6FKRRORI%LRVWXGLHV.\RWR8QLYHUVLW\(LLFKLUR6X]XNL

7DNHKLNR.RED\DVKL-XQLFKL1DND\DPD([HFXWLYH'LUHFWRU.LKDUD0HPRULDO<RNRKDPD)RXQGDWLRQ

3URIHVVRU,QVWLWXWHIRU4XDQWLWDWLYH%LRVFLHQFHV7RN\R8QLYHUVLW\

3URIHVVRU1DWLRQDO,QVWLWXWHIRU%DVLF%LRORJ\1DWLRQDO,QVWLWXWHVRI1DWXUDO6FLHQFHV

0RWRPX(QGR

0LNL6KLPDGD

3URIHVVRU'LYLVLRQRI%LRORJLFDO6FLHQFH1DUD,QVWLWXWHRI6FLHQFHDQG7HFKQRORJ\
3URIHVVRU+HDGRI'LYLVLRQRI3KDUPDF\7RWWRUL8QLYHUVLW\+RVSLWDO.RLFKL)XNDVH

3URIHVVRU*UDGXDWH6FKRRORI6FLHQFH2VDND8QLYHUVLW\

6KLQML6KLPRMR

3URIHVVRU&\EHUPHGLD&HQWHU2VDND8QLYHULVLW\

7DNDVKL1DJDVDZD

3URIHVVRU*UDGXDWH6FKRRORI)URQWLHU%LRVFLHQFHV2VDND8QLYHUVLW\.D]X\D.LNXFKL

3URIHVVRU*UDGXDWH6FKRRORI(QJLQHHULQJ2VDND8QLYHUVLW\+LURDNL0LNL

3URIHVVRU5HVHDUFK,QVWLWXWHIRU0LFURELDO'LVHDVHV◦◦◦ 65 ◦IPR
6WDIIDQG$FFRXQWLQJ'DWD

$VRI0D\

6WDII

9LVLWLQJ&ROODERUDWLYH5HVHDUFKHUV$VVRFLDWH3URIHVVRU$FDGHPLF6WDII

5HVHDUFKHU$VVRFLDWH3URIHVVRU/HFWXUHU$VVLVWDQW3URIHVVRU

3URIHVVRURI'RQDWHG)XQG5HVHDUFK



7RWDO



$VVRFLDWH3URIHVVRURI'RQDWHG)XQG5HVHDUFK,QWHUQDWLRQDO-RLQW5HVHDUFK&ROODERUDWRU$VVLVWDQW3URIHVVRURI'RQDWHG)XQG5HVHDUFK-RLQW5HVHDUFK&ROODERUDWRU

6SHFLDOO\DSSRLQWHG3URIHVVRU)XOO7LPH 6SHFLDOO\DSSRLQWHG$VVRFLDWH3URIHVVRU



6SHFLDOO\DSSRLQWHG/HFWXUHU

6SHFLDOO\DSSRLQWHG$VVLVWDQW3URIHVVRU



6SHFLDOO\DSSRLQWHG5HVHDUFKHU7HFKQLFDO6WDII

6SHFLDOO\DSSRLQWHG7HFKQLFDO6WDII$GPLQLVWUDWLYH6WDII

6SHFLDOO\DSSRLQWHG$GPLQLVWUDWLYH6WDII

6XEWRWDO

9LVLWLQJ
5HVHDUFKHU-RLQW

5HJXODU

5HVHDUFK%HDPOLQH &ROODERUDWRUWRWDOLQ

105



&U\R(0



7RWDO





6WXGHQWV

8QGHUJUDGXDWH6SHFLDOO\DSSRLQWHG5HVHDUFKHU*UDGXDWH6WXGHQWʤ0DVWHU&RXUVHʥ7HFKQLFDO6WDII*UDGXDWH6WXGHQWʤ'RFWRU&RXUVHʥ

3DUW7LPH $GPLQLVWUDWLYH6WDII

5HVHDUFK6WXGHQW&RPPLVVLRQHG6WDII

6XEWRWDO
7RWDO



3URIHVVRU

3URIHVVRU$FFRXQWLQJLQ)<  ◦ 66 ◦◦◦ About IPR







7RWDO



Institute for Protein Research Osaka University)ORRU3ODQ
Structural Analysis Research Building

Main Building

9F

ɻɻ5HVHDUFK&HQWHUIRU6WDWHRIWKH$UWɻɻɻɻ)XQFWLRQDO3URWHLQ$QDO\VLV$QLPDO)DFLOLWLHV$QLPDO)DFLOLWLHV8F+LNLGDODE

7F

4F

5DGLR,VRWRSH)DFLOLWLHV)858.$:$ODE/DERI3URWHLQ'DWDEDVHV

3F

6+,12+$5$
ODE

6HPLQDUURRP

1$.$*$:$
ODE

2F+$5$'$ODE

2.$'$ODE

1$*$12ODE

6F;UD\IDFLOLW\

1F*272ODE

5F&RPPRQIDFLOLWLHV

1$.$,ODE

Analytical Instruments Facility
7$.$*,ODE)8-,:$5$ODE

/DERI$GYDQFHG3URWHLQ&KDUDFWHUL]DWLRQ$GPLQLVWUDWLRQ/DUJH
0HGLXP
2⒑FH
PHHWLQJ 'LUHFWRUbV PHHWLQJURRPTAKAO lab

Library
6ROXWLRQ
105

URRP

/RXQJH

6ROLGVWDWH
105

Lecture Hall

Building Layout

Collaborative Research Facility&ROODERUDWLYH
5HVHDUFK)DFLOLW\
105
)DFLOLW\/DERI$GYDQFHG3URWHLQ&KDUDFWHUL]DWLRQ

1F/DERI0XOWLVFDOH6WUXFWXUDO%LRORJ\

1F

NMR Group+RMRODE

0LPDODEURRP

EM Group.85,68ODE

3F

2F

2F

6HPLQDUURRP

4F

NMR Facility$QDO\VLV,QVWUXPHQWV)DFLOLW\

2F

2SHQVSDFHODE
0DWUL[RPHVWHPFHOOYHQWXUH<RXQJ,QGHSHQGHQW5HVHDFKHUVODE

0DLQ%XLOGLQJ.$12+ODE

6WUXFWXUDO$QDO\VLV
5HVHDUFK%XLOGLQJ'LYLVLRQRI0DWUL[RPH
5HVHDUFKDQG$SSOLFDWLRQ

1F◦◦◦ 67 ◦IPR
ACCESS MAP◦ 68 ◦◦◦ ACCESS MAPIPRInstitute for Protein Research Osaka University

CONTENTS
Message from the Director

1

Concept and Future2

Organization3

Sixty-year history of IPR

4

Activities 2017-2018

6

Division of Protein Chemistry

9

Laboratory of Protein Organic Chemistry
Laboratory of Cell Systems
Laboratory of Nanobiology
Laboratory of Organelle Biology

Division of Protein Structural Biology

17

Laboratory of Protein Folding
Laboratory of Molecular Biophysics
Laboratory of Protein Crystallography
Laboratory of Membrane Protein Chemistry

Division of Integrated Protein Functions

25

Laboratory of Genome and Chromosome Functions
Laboratory for Molecular and Developmental Biology
Laboratory for Advanced Brain Functions
Laboratory of Homeostatic Integration

Division of Multiscale Integrative Protein Science

33

Laboratory of Nuclear Network
Laboratory of Multiscale Structural Biology
Laboratory for Infection Systems
Laboratory of Visiting Scientists

Research Center for State-of-the-Art Functional Protein Analysis

39

Laboratory of Protein Profiling and Functional Proteomics
Laboratory of Protein Synthesis and Expression
Laboratory of Supramolecular Crystallography
Laboratory of Advanced Protein Characterization
Laboratory of Protein Databases
Open Space Laboratory for Advanced Protein Science

Technology Division

53

Technology Division

Division of Donated Fund Research

55

Division of Matrixome Research and Application

About IPR

57

Educational Activities
Research Activities

Public Relations Office

Joint-Usage / Research Center
IPR Saff

Institute for Protein Research, Osaka University

Administrative Council

3−2 Yamadaoka, Suita Osaka, 5650871, Japan

Staff and Accounting Data

http://www.protein.osaka-u.ac.jp

Floor PlanACCESS MAP

68

3Prospectus 2018 Institute for Protein Research Osaka University

OSAKA UNIVERSITY

IPR
Prospectus 2018

Institute
I tit t for
f Protein
P t i ResearchRhOsaka University