4th RSC-CSJ
Joint Symposium 2013
Chemical Biology Researchby Young Investigators

March 24, 2013, the 93rd CSJ Annual Meeting
Ritsumeikan University
Biwako-Kusatsu Campus, Shiga, JAPANRoyal Society of Chemistry & Chemical Society of Japan

4th RSC-CSJ表紙案.indd 1

13/02/19 23:25

4th RSC－CSJ Joint Symposium 2013
―Chemical Biology Research by Young Investigators―
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Date:
Venue:March 24th (SUN) 9:00‐17:40
Room S8 (CO‐LEARNING HOUSE I C306),
Ritsumeikan University, Biwako‐kusatsu Campus(Shiga, Japan)
Hosted by The Chemical Society of Japan
Co‐hosted by Royal Society of Chemistry□Chair: Kazuya Kikuchi
09:10‐09:20
Opening Remarks from CSJ
09:20‐09:30
Opening Remarks from UK□Chair: Hiroshi Murakami
L‐1
09:30‐10:10
Gregory L. CHALLIS(University of Warwick)“Elucidating and Exploiting Novel Biosynthetic CH Functionalization Reactions”

page 1□Chair: Gregory L. Challis
L‐2
10:10‐10:50
Hiroshi MURAKAMI(The University of Tokyo) page 3“Development of Non‐standard Peptide Inhibitors Using TRAP Display”
10:50‐11:20

Intermission□Chair: Kazushi Kinbara
L3
11:20‐12:00
Dominic J. CAMPOPIANO (University of Edinburgh) page 5“Sphingolipid Biosynthesis in Man and Microbes”□Chair: Dominic Campopiano
L‐4
12:00‐12:40
Kazushi KINBARA(Tohoku University)“Development of Supramolecular Tools for Manipulation of Biological Molecules”
12:40‐13:40

page 7

Lunch Time

□Chair: Stuart J. Conway
L‐5
13:40‐14:20
Motonari UESUGI(Kyoto University)“Small Molecule Tools for Cell Biology and Cell Therapy”

page 9□Chair: Motonari Uesugi
page 11
L‐6
14:20‐15:00
Stuart J. CONWAY(University of Oxford)“Cracking the Histone Code: Inhibitors of the Bromodomain‐AcetylLysine Interaction”
15:00‐15:30

Intermission□Chair: Rebecca GOSS
L‐7
15:30‐16:10
Masayuki INOUE(The University of Tokyo) page 13“Total Synthesis and Biological Evaluation of Polytheonamide B”□Chair: Masayuki Inoue
L‐8
16:10‐16:50
Rebecca GOSS
“Elucidating and Exploiting Biosynthesis”(University of St Andrews) page 15□Chair: Rebecca Goss
L‐9
16:50‐17:30
Kazuya KIKUCHI(Osaka University)"Design, Synthesis and Biological Application of in Vivo Imaging Probes with
Tunable Chemical Switches”
17:30‐17:40

Closing Remarks

page 17

Welcome Address
Dear Colleagues,
Welcome to the 4th RSC-CSJ Joint Symposium 2013
―Chemical Biology Research by Young Investigators―, to beheld on March 24, 2013 at the same venue as the CSJ 93rd
Annual Meeting; Ritsumeikan University, Biwako-Kusatsu
Campus, Shiga, located by the shore of Lake Biwa, under theco-sponsorship of the Chemical Society of Japan (CSJ) and theRoyal Society of Chemistry (RSC). This series of bilateral
symposium originates from the first one in 2007 held in Osaka
University, followed by the second in 2008 at Queen’s University
and the third in 2010 at the RSC Burlington House where we
signed an International Cooperation Agreement between RSC
and CSJ.
There are two features to be mentioned for the present symposium.
Firstly, this 4th Joint Symposium on Chemical Biology Research was originally planned
to be held at the CSJ 91st Annual Meeting late March 2011 at Kanagawa University, but
unfortunately it was forced to be cancelled by the March 11 Disasters, earthquakes andnuclear power plant accidents. The original plan has now revived to be held at this 93rd
Annual Meeting after one-year blank. I would like to thank the symposium chair Prof.
Kazuya Kikuchi, and RSC Chief Executive Officer Dr. Robert Parker as well as CSJ
Executive Director Mr. Nobuyuki Kawashima, and also Dr. Hirofumi Seike, Representativeat the RSC Tokyo Office, for their great efforts to make this symposium possible.
Secondly, this year 2013 marks The 150 Year Anniversary of UK-Japan Academic
Interaction, since five young nobles from Choshu clan, so-called Chosu-Five, including Ito
Hirobumi and Inoue Kaoru, visited UK in 1863 to study at University College London(UCL). Furthermore, 19 students from Satsuma clan visited UCL in 1865 and Joji Sakurai,
now known as the Father of Chemistry in Japan, became a student at UCL in 1876. They
were all nurtured mainly by the famous chemist Prof. Alexander Williamson, President of
the London Chemical Society (a predecessor of RSC). So, it is a great pleasure for us to holdthe UK/Japan Joint Symposium in this special year, by thanking all the young talents fromJapan and many scientists from UK for their pioneering contribution to establish the strong
bridge between two countries UK and Japan, and between RSC and CSJ. I really hope that
this joint symposium will further strengthen the bridge for our scientific cooperation.
For this symposium nine distinguished chemists in the area of chemical biology, four
from UK and five from Japan, will present their research results and exchange ideas forfurther development. Chemical biology is a basic science for both “life innovation” and“green innovation”, which have been claimed as the science and technology policy forcreation of sustainable society. I really hope that this bilateral symposium will contribute
greatly not only to the rapidly developing chemical biology field but also a lot of related
fields. It is also my great expectation that all of the attendants strengthen your friendshipand find new friends throughout the symposium for further collaborations.
Kohei Tamao

President
The Chemical Society of Japan

Welcome Address
Dear colleagues,
The Royal Society of Chemistry (RSC) isdelighted to be co-organising with the Chemical Society ofJapan (CSJ) this RSC-CSJ Joint Symposium on Chemical
Biology. We would like to thank Professors Kazuya Kikuchi
and Nobuyuki Kawashima for all their help and support tomake this event possible. This symposium is one of theactivities jointly organised following the signing in 2010 of
an International Cooperation Agreement between the RSC
and the CSJ. We hope it will be another successful andmemorable CSJ-RSC activity that will facilitate and foster
future collaborations between the societies and scientists in both countries.
The RSC is a learned society, concerned with advancing the chemical sciences,developing its applications, and disseminating chemical knowledge. It is also a
professional body that maintains professional qualifications and sets high standards ofcompetence and conduct for professional chemists. We have 48,000 members
worldwide drawn from all areas of the chemical sciences, including chemical biology.
Our Chemical Biology Interface Division aims to improve interdisciplinary research at
the chemistry-biology interface, and promote the importance of this area to thegovernment, industry, academia and education. In addition to developing young talents
and advancing the chemical sciences both in the UK and internationally, the RSC is amajor publisher of research journals, magazines, databases and books that cover all
areas of the chemical sciences.
The RSC is also pleased to have the opportunity to work more closely with
the Japanese chemical science community through Dr Hirofumi Seike, ourRepresentative for Japan, who is based full-time in our office in Tokyo. This office islocated in the CSJ building in Tokyo and we are very grateful to the CSJ for all their
help and advice in establishing this office. Dr. Seike is happy to answer your queries
about any aspects of the RSC including its activities in Japan, its membership services
and publishing activities.
Each of the speakers in this symposium is an international expert in the areaof chemical biology, which underpins the development of new medicine and theunderstanding of biological processes. We hope that the presentations will stimulate
the exchange of ideas and experiences between all participants. We thank each of the
speakers and all the participants for their contributions to this symposium.
I offer a very warm welcome to what promises to be an exciting scientificevent. We hope that the symposium will provide a springboard for future activities andthat it will foster new research collaborations. We look forward to continuing our close
partnership with CSJ, with the possibility of more joint CSJ-RSC activities in the future.

Dr Robert Parker
Chief Executive Officer
Royal Society of ChemistryGregory CHALLIS
Department of Chemistry,
University of Warwick

Educational Background B. Sc. 1994, Imperial College London
 Ph. D. 1998, the University of Oxford(advisor: Sir Jack Baldwin FRS) Postdoctoral Fellow, 1998-2000, Johns Hopkins University(advisor: Craig Townsend) Postdoctoral Fellow, 2000-2001, John Innes Centre(advisor: Keith Chater FRS ) Lecturer (Assistant Professor), 2001-2003, University of Warwick Senior Lecturer (Associate Professor), 2003-2006, University of Warwick
 Professor (2006)-present, University of WarwickScientific Interests
Natural products chemistry and biology, including isolation and structure determination of
new bioactive natural products, genomics and genetics of natural product biosynthesis,
enzymology of natural product biosynthesis, chemical synthesis of bioactive natural products
and intermediates in their biosynthesis, genetic manipulation of bioactive natural productbiosynthesis pathways to produce new analogues, molecular mechanism of action of
bioactive natural products and their biological function. A highly multidisciplinary approachto these problems is taken encompassing high field NMR spectoscopy, mass spectrometry,
HPLC, organic synthesis, bioinformatics, microbiological methods, molecular genetic
manipulation, recombinant protein overproduction and purification, and enzyme assay
development.

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Recent Papers
1. “Cytochrome P450–Catalyzed L-Tryptophan Nitration in Thaxtomin Phytotoxin
Biosynthesis” S. M. Barry, J. A. Kers, E. G. Johnson, L. Song, P. R. Aston, B. Patel, S. B.
Krasnoff, B. R. Crane, D. M. Gibson, R. Loria, G. L. Challis, Nat. Chem. Biol. 2012, 8, 814816.
2. “Regio- and Stereodivergent Antibiotic Oxidative Carbocyclizations Catalyzed by Rieske
Oxygenase-Like Enzymes” P. K. Sydor, S. M. Barry, O. M. Odulate, F. Barona-Gomez, S. W.
Haynes, C. Corre, L. Song, G. L. Challis, Nat. Chem. 2011, 3, 388-392.
3. “Identification of a Bioactive 51-Membered Macrolide Complex by Activation of a Silent
Polyketide Synthase in Streptomyces ambofaciens” L. Laureti, L. Song, S. Huang, C. Corre, P.
Leblond, G. L. Challis, B. Aigle, Proc. Natl. Acad. Sci. USA, 2011, 108, 6258-6263.
4. “AcsD Catalyzes Enantioselective Citrate Desymmetrization in Siderophore Biosynthesis” S.
Schmelz, N. Kadi, S. A. McMahon, L. Song, D. Oves-Costales, M. Oke, H. Liu, K. A.
Johnson, L. G. Carter, C. H. Botting, M. F. White, G. L. Challis, J. H. Naismith, Nat. Chem.
Biol. 2009, 5, 174-182.
5. “2-Alkyl-4-hydroxymethylfuran-3-carboxylic Acids, Antibiotic Production Inducers
Discovered by Streptomyces coelicolor Genome Mining” C. Corre, L. Song, S. O'Rourke, K.
F. Chater, G. L. Challis, Proc. Natl. Acad. Sci. USA. 2008, 105, 17510-17515.
6. “A New Family of ATP-dependent OligomerizationMacrocyclization Biocatalysts” N. Kadi,
D. Oves-Costales, F. Barona-Gomez, G. L. Challis, Nat. Chem. Biol. 2007, 3, 652-656.
7. “Discovery of a New Peptide Natural Product by Streptomyces coelicolor Genome Mining” S.
Lautru, R. J. Deeth, L. M. Bailey, G. L. Challis. Nat. Chem. Biol. 2005, 1, 265-269.
4th RSC－CSJ Joint Symposium 2013
―Chemical Biology Research by Young Investigators―1

L-1
Elucidating and Exploiting Novel Biosynthetic CH
Functionalization Reactions
Gregory L. CHALLIS
Department of Chemistry, University of Warwick, Coventry CV4 7AL, UK
g.l.challis@warwick.ac.uk

CH functionalization reactions are key steps in the biosynthesis of numerous bioactive
natural products. Such reactions include specific hydroxylation, chlorination and desaturation ofunactivated carbon atoms, as well as a variety of oxidative cyclization reactions, exemplified by
the conversion of the tripeptide ACV to the bicyclic penicillin nucleus.
Here we report two unprecedented types of enzyme-catalyzed CH functionalization
reaction in natural product biosynthesis. RedG and McpG, novel Rieske non-heme iron dependentoxygenase-like enzymes, catalyse the regio- and stereodivergent oxidative carbocyclization ofundecylprodigiosin to streptorubin B and metacycloprodigiosin, respectively, and TxtE, a unique
cytochrome P450, catalyzes regiospecific nitration of L-tryptophan, the first committed step inthaxtomin phytotoxin biosynthesis. Efforts to elucidate the catalytic mechanisms of these enzymes
and to exploit them for the production of novel streptorubin B and nitrotryptophan derivatives will
be described.

2

The 93rd CSJ Annual Meeting
24th March 2013, Ritsumeikan University, Shiga

Hiroshi MURAKAMI
Department of Life Sciences,
Graduate School of Arts and Sciences,
The University of Tokyo Educational Background
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B.S. 1995, Okayama University
Ph.D. 2000, Okayama University (Prof. Masahiko Sisido)
Postdoctoral Fellow, 2000-2001, Okayama University
Postdoctoral Fellow, 2001-2003, State University of New York at Buffalo(Prof. Hiroaki Suga)
Assistant Professor, 2003-2009 The University of Tokyo (Prof. Hiroaki Suga)
Associate Professor, 2009-, The University of Tokyo (Principal Investigator)

 Scientific Interests
Chemical Biology and Biomolecular Engineering: Directed evolution of new functional
biomolecules that are useful for biological and therapeutic applications.

 Recent Papers
1. “Reevaluation of the D-Amino Acid Compatibility with the Elongation Event inTranslation” Fujino, T.; Goto, Y.; Suga, H.; Murakami, H.*, J. Am. Chem. Soc. 2013, 135,(5), 1830-7
2. “Genetically Encoded Libraries of Nonstandard Peptides” Kawakami, T.; Murakami, H. *,J. Nucl. Acids 2012, 713510.
3. “Bases in the Anticodon Loop of tRNA(Ala)(GGC) Prevent Misreading” Murakami, H.*;
Ohta, A.; Suga, H.*, Nat. Struct. Mol. Biol. 2009, 16, (4), 353-8.
4. “Diverse Backbone-cyclized Peptides via Codon Reprogramming” Kawakami, T.; Ohta,
A.; Ohuchi, M.; Ashigai, H.; Murakami, H.; Suga, H.*, Nat. Chem. Biol. 2009, 5, (12),888-90.
5. “Structural Basis of Specific tRNA Aminoacylation by a Small In Vitro Selected
Ribozyme” Xiao, H.; Murakami, H.; Suga, H.; Ferre-D'Amare, A. R.*, Nature 2008, 454,
(7202), 358-61.
6. “Ribosomal Synthesis of Bicyclic Peptides via Two Orthogonal Inter-side-chain
Reactions” Sako, Y.; Morimoto, J.; Murakami, H.; Suga, H.*, J. Am. Chem. Soc. 2008, 130,(23), 7232-4.
7. “Messenger RNA-programmed Incorporation of Multiple N-Methyl-amino Acids into
Linear and Cyclic Peptides” Kawakami, T.; Murakami, H.; Suga, H.*, Chem. Biol. 2008,
15, (1), 32-42.
8. “Reprogramming the Translation Initiation for the Synthesis of Physiologically Stable
Cyclic Peptides” Goto, Y.; Ohta, A.; Sako, Y.; Yamagishi, Y.; Murakami, H.; Suga, H.*,ACS Chem. Biol. 2008, 3, (2), 120.
9. “A Highly Flexible tRNA Acylation Method for Non-natural Polypeptide
Synthesis ”Murakami, H.; Ohta, A.; Ashigai, H.; Suga, H.*, Nat. Methods 20 06, 3, (5),357-9.

4th RSC－CSJ Joint Symposium 2013
―Chemical Biology Research by Young Investigators―3

L-2
Development of Non-standard Peptide Inhibitors
Using TRAP Display
Hiroshi MURAKAMI
Department of Life Sciences, Graduate School of Arts and Sciences,
The University of Tokyo, 3-8-1, Komaba, Meguro-ku, Tokyo 153-8902, Japanmurah@bio.c.u-tokyo.ac.jp
In vitro selection using a peptide library linked with its genotype (DNA/RNA) are powerfultools to discover novel functional peptides. Compared with high-throughput screening ofchemically synthesized peptide libraries, the selection methods can deal with much larger libraries
of up to 1013 peptides, which dramatically increases the probability of getting peptides withdesired properties.
To apply this methodology for a non-standard peptide that contains non-proteinogenic
structure for stabilizing the peptide in in vivo, we developed a cell-free translation system whose
genetic code was reprogrammed to introduce non-proteinogenic amino acids. Using the translation
system, we studied the compatibility of many types of non-proteinogenic amino acids including Nmethyl amino acids, D-amino acids, β-amino acids with the translation. We found that thecompatibility of these amino acids was strongly depended on the structure of the side chain. For
example, D-amino acids with non-bulky alkyl or aromatic side chains (Ala, Ser, Cys, Met, Thr, His,
Phe, and Tyr) showed the incorporation efficiencies comparable with the corresponding L-isomer,
and these with uncharged hydrophilic or alkyl side chains (Asn, Gln, Val, and Leu) showeddecreased incorporation efficiencies. The D-amino acids with other property (Arg, Lys, Asp, Glu,
Ile, Trp, and Pro) were not incorporated into a peptide.
Concurrently with the above screening of non-proteinogenic amino acids, we also
developed a new in vitro display method, TRAP (Transcription–translation coupled with
Association of Puromycin-linker) display method, that enables high-speed in vitro selection offunctional peptides, peptidomimetics via a simple procedure. Using TRAP display, macrocyclic
non-standard peptides that bind to human serum albumin were selected. Six rounds of selection
using TRAP display were performed only in approximately 14 h. We have also applied TRAP
display for the in vitro selection of macrocyclic non-standard peptides against vascular endothelial
growth factor receptor 2 (VEGFR2). The eight macrocyclic-peptide libraries consisted of
combinations of four variants of amino acid linkers for cyclization and two versions of elongatoramino acid compositions, including four backbone-modified non-proteinogenic amino acids.
Affinity selection from these libraries, yielded multiple anti-VEGFR2 macrocyclic peptide leads.
By further screening of the chemically synthesized lead peptides for antagonizing activity, we
identified a potent macrocyclic peptide that inhibited VEGF-induced VEGFR2
autophosphorylation, proliferation and angiogenesis of vascular endothelial cells.
Through the combination of the codon reassignment and the TRAP display enables highspeed selection of functional non-standard peptides, it will facilitate the generation of various new
non-standard peptides that are useful for biological and therapeutic applications.

4

The 93rd CSJ Annual Meeting
24th March 2013, Ritsumeikan University, Shiga

Dominic CAMPOPIANO
School of Chemistry, EastChem,
University of EdinburghWest Mains Road, Edinburgh, EH9 3JJEducational Background
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B. Sc. 1988, University of Glasgow
Ph. D. 1994, University of Edinburgh(advisor: Robert L. Baxter)
Postdoctoral Fellow, 1992-1995, University of Leicester, UK(advisor: William V. Shaw)
Postdoctoral Fellow, 1995-1996, University of Edinburgh, UK(advisor: Robert Baxter)
Lecturer 1997-2007, University of Edinburgh
Senior Lecturer, 2007-present, University of Edinburgh
Royal Society of Edinburgh/Scottish Executive Research Fellow, 2006.

Scientific Interests
Chemical Biology: Natural product biosynthesis, enzyme mechanism, protein structure, host-pathogen
interactions, dynamic covalent chemistry, biocatalysis, directed evolution.

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Recent Papers
1.
2.
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7.
8.

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“An Improved Racemase/Acylase Biotransformation for the Preparation of Enantiomerically
Pure Amino Acids” Baxter, S., Royer, S., Grogan, G., Brown, F., Holt-Tiffin, K. E., Taylor, I.
N., Fotheringham, I. G. , Campopiano, D. J., J. Am. Chem. Soc. 2012, 134, 19310-19313.
“Role of a Conserved Arginine Residue during Catalysis in Serine Palmitoyltransferase"
Lowther, J., Charmier, G., Raman, M. C., Ikushiro, H., Hayashi, H., Campopiano, D. J., FEBS
Lett 2011, 585, 1729-1734.
“Structural, Mechanistic and Regulatory Studies of Serine Palmitoyltransferase” Lowther, J.,
Naismith, J. H., Dunn, T. M., Campopiano, D. J., Biochem. Soc. Trans. 2012, 40, 547-554.
“The Serine Palmitoyltranserase from Sphingomonas wittichii RW1: An Interesting Link to an
Unusual Acyl Carrier Protein” Raman, M. C. C., Johnson, K. A., Clarke, D. J., Naismtih, J. H.,
Campopiano, D. J. Biopolymers, Special Issue on Natural Products 2010, 93, 811-822.
“Serine Palmitoyltranserase Displays a Novel Mechanism of Cycloserine Inhibition” Lowther,J., McMahon, S., Johnson, K. A., Yard, B. A., Carter, L. G Raman, M. C., Naismith, J. H.,
Campopiano, D. J., Molecular Biosystems, Emerging Investigators, Special Issue 2010, 6,
1682-1693.
“Nucleophilic Catalysis of Acylhydrazone Equilibration for Protein-directed Dynamic CovalentChemistry” Bhat, V. T., Caniard, A. M., Luksch, T., Brenk, R., Campopiano, D. J.,* Greaney,
M. F.*, Nat. Chem. 2010,2, 490-497.
“Insights into How Nucleotide-binding Domains Power ABC Transport” Newstead, S., Fowler,
P. W., Bilton, P., Carpenter, E. P., Sadler, P. J., Campopiano, D. J.*, Sansom, M. S. P.*, Iwata,
S.*, Structure 2009, 17, 1213-1222.
“The External-aldimine form Of Serine Palmitoyltranserase; Structural, Kinetic, andSpectroscopic Analysis of athe Wild-Type Enzyme and HSAN1 Mutant Mimics” Raman, M. C.,
Johnson, K. A., Yard, B. A., Lowther, J., Carter, L. G., Naismith, J. H.*, Campopiano, D. J.*, J.
Biol. Chem. 2009, 284, 17328-17339.
“The Structure of Serine Palmitoyltransferase; Gateway to Sphingolipid Biosynthesis” Yard, B.
A., Carter, L. G., Johnson, K. A., McMahon, S., Dorward, M., Liu, H., Oke, M., Overton, I.,
Barton, G. J., Naismith, J. H., Campopiano, D. J., J. Mol. Biol. 2007, 370, 870-886.

4th RSC－CSJ Joint Symposium 2013
―Chemical Biology Research by Young Investigators―

5

L-3
Sphingolipid Biosynthesis in Man and Microbes
Dominic J. CAMPOPIANO
School of Chemistry, EastChem, University of Edinburgh, West Mains Road,
Edinburgh, EH9 3JJ, UK
Dominic.Campopiano@ed.ac.uk
Sphingolipids (SLs) are essential components of all eukaryotic and many prokaryotic membranes
where they play essential structural roles. SLs are long chain bases (lcbs) composed of fatty acids and apolar head group (derived from L-serine). In recent years SLs have also been shown to act as potent
signaling molecules that participate in various cellular functions. For example, sphingosine 1phosphate is a potent signal molecule that it known to have five G-protein coupled receptors (GPCRs)
in the nuclear membrane. Errors in sphingolipid metabolism have been implicated in a number of
inflammatory diseases and cancer. Recent efforts by a large consortium (Lipidmaps) have sought toinventory the large, diverse and chemically-complex family of SLs using high resolution structural
techniques such as mass spectrometry. With this catalogue in hand it is now possible to study theregulation of SL biosynthesis in healthy and diseased cells and provide a systems biology roadmap of
SL metabolism. It is hoped that our understanding may lead to the development of therapeutic agents
specifically directed at targets from SL pathways.
We have been studying SL biosynthesis with a goal to increase our understanding of how eachenzyme in the pathway catalyses a particular reaction and how it is regulated. Later stages of SL
biosynthesis are species-specific but all core SLs are synthesized from the condensation of L-serine
and a long-chain fatty acid thioester such as palmitoyl-CoA (C16). This is catalysed by the enzyme
serine palmitoyl transferase (SPT). SPT is a pyridoxal 5’ phosphate (PLP)-dependent enzyme thatcatalyses the formation of the alpha-oxoamine product, 3-ketodihydrosphingosine (KDS) through adecarboxylative, Claisen-like condensation reaction. Eukaryotic SPTs are membrane-bound,heterodimeric enzymes encoded by two genes (lcb1, lcb2) whereas bacterial enzymes are cytoplasmic
homodimers encoded by a single gene. Due to the technical problems associated with studying the
mammalian enzymes we have used bacterial SPTs (from Sphingomonas strains) as models to study
their structure and mechanism using various spectroscopic methods including x-ray crystallography.
We found the PLP-binding site at the dimer interface of the bacterial SPT homodimer. Mutations inhuman SPT cause hereditary sensory autonomic neuropathy type 1 (HSAN), a rare diseasecharacterized by loss of feeling in extremities and severe pain. The molecular basis of how thesemutations perturb SPT activity is subtle and is not simply loss of activity. It is thought that the mutant
SPT is promiscuous and generates toxic sphingolipids. We used our bacterial SPT structure to mimic
the human HSAN1 mutants and found that mutations on one subunit impact on the enzyme activity
and structure of the dimer. We have also explored the nature of SPT inhibition using both enantiomersof the drug cycloserine. We have begun to investigate the complete biosynthetic pathway using various
Sphingomonas species as model organisms. We are using knowledge gained from these studies toprobe the more complex mammalian systems.

6

The 93rd CSJ Annual Meeting
24th March 2013, Ritsumeikan University, Shiga

Kazushi KINBARA
Institute of Multidisciplinary Research for Advanced Materials,
Tohoku University
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Educational Background
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Scientific Interests
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B. S. 1991, University of Tokyo
Ph. D. 1996, University of Tokyo (advisor: Kazuhiko Saigo)
Research Associate, 1996-2001, The University of Tokyo
Lecturer, 2001-2006, The University of Tokyo
Associate Professor, 2006-2008, The University of Tokyo
Professor, 2008-Present, Tohoku University

Development of biomimetic molecules
Supramolecular chemistry of macromolecules
Protein engineering

Recent Papers
1. “Ion Permeation by a Folded Multiblock Amphiphilic Oligomer Achieved by HierarchicalConstruction of Self-assembled Nanopores” T. Muraoka, T. Shima, T. Hamada, M. Morita, M.
Takagi, K. V. Tabata, H. Noji, K. Kinbara, J. Am. Chem. Soc. 2012, 134, 19788–19794.
2. “Application of Photoactive Yellow Protein as a Photoresponsive Module for Controlling Hemolytic
Activity of Staphylococcal -Hemolysin” M. Ui, Y. Tanaka, Y. Araki, T. Wada, T. Takei, K.
Tsumoto, K. Kinbara, Chem. Commun. 2012, 48, 4737–4739.
3. “Mimicking Multipass Transmembrane Proteins: Synthesis, Assembly and Folding of Alternating
Amphiphilic Multiblock Molecules in Liposomal Membranes” T. Muraoka, T. Shima, T. Hamada,
M. Morita, M. Takagi, K. Kinbara, Chem. Commun. 2011, 47, 194–196.
4. “Adhesion Effects of a Guanidinium Ion Appended Dendritic ‘Molecular Glue’ on the ATP-driven
Sliding Motion of Actomyosin” K. Okuro, K. Kinbara, K. Takeda, Y. Inoue, A. Ishijima, T. Aida,
Angew. Chem., Int. Ed. 2010, 49, 3030–3033.
5. “Shape-directed Assembly of a ‘Macromolecular Barb’ into Nanofibers: StereospecificCyclopolymerization of Isopropylidene Diallylmalonate” Y. Miyamura, K. Kinbara, Y. Yamamoto,
V. K. Praveen, K. Kato, M. Takata, A. Takano, Y. Matsushita, E. Lee, M. Lee, T. Aida, J. Am. Chem.
Soc. 2010, 132, 3292–3294.
6. “A Tubular Biocontainer: Metal Ion-induced 1D Assembly of a Molecularly Engineered
Chaperonin” S. Biswas, K. Kinbara, N. Oya, N. Ishii, H. Taguchi, T. Aida, J. Am. Chem. Soc. 2009,
131, 7556–7557.
7. “Molecular Glues Carrying Multiple Guanidinium Ion Pendants via an Oligoether Spacer:
Stabilization of Microtubules against Depolymerization” K. Okuro, K. Kinbara, K. Tsumoto, N. Ishii,
T. Aida, J. Am. Chem. Soc. 2009, 131, 1626–1627.
8. “Toward Long-distance Mechanical Communication: Studies on a Ternary Complex Interconnectedby a Bridging Rotary Module” H. Kai, S. Nara, K. Kinbara, T. Aida, J. Am. Chem. Soc. 2008, 130,
6725–6727.

4th RSC－CSJ Joint Symposium 2013
―Chemical Biology Research by Young Investigators―

7

L-4
Development of Supramolecular Tools
for Manipulation of Biological Molecules
Kazushi KINBARA
Institute of Multidisciplinary Research for Advanced Materials, Tohoku
University
2-1-1, Katahira, Aoba-ku, Sendai, Miyagi 980-8577, Japankinbara@tagen.tohoku.ac.jp
Engineered macromolecules capable of interfacing biological events have become more
important in the field of nanobiotechnology. In particular, development of molecular tools that areable to control function of proteins is a hot issue in both materials science and nanomedicine. For
example, polymerization/depolymerization event of microtubules, tubular assemblies of  −tubulin heterodimers, play essential roles in mitosis. In connection with this, paclitaxel, which
stabilizes microtubules against depolymerization, has been applied for anticancer drugs.
We have developed ‘molecular glue’ which is capable of non-covalent attachment to various
wild-type proteins via a multivalent salt-bridge formation between guanidinium (Gu+) ions andoxyanionic groups that exist ubiquitously in proteins. Gu+ is known to strongly associate withseveral oxyanion functionalities such as carboxylate and phosphate groups by electrostatic andhydrogen bonding interactions. Various biomolecules such as proteins, lipids, and nucleic acids
have anionic functionalities, and therefore can be hybridized with Gu+ groups. Actually, we havefound that the molecular glues efficiently stabilize protein assemblies such as microtubules (MTs)and actomyosin, and prevent them from dissociation into subunits.
On the other hand, we have focused on polyethylene glycol (PEG), which is a linear polymer
widely used for industrial materials to medicines.

Nonionic, water-soluble and nontoxic

characters of PEG are particularly useful for pharmaceutical applications. Covalent attachment of
PEG chains to a target molecule such as a drug or a therapeutic protein, is know as “PEGylation”,
that inhibits protein aggregation and prolongation of drugs’ circulatory time. However, it is also
known that linear PEG chains tend to cover the surface of proteins to cause serious deactivation of
their functions. Here we propose novel molecules composed of ethylene oxide skeleton, as“structuralized” PEG molecules, that adopt rigid 2D or 3D structures while holding the merits of
PEG. As the first target, we designed triangle molecules composed of tetraethylene glycol as thelinker unit and pentaerythritol as the vertex unit, and found that they have characteristic properties
compared with the linear PEG molecules.

8

The 93rd CSJ Annual Meeting
24th March 2013, Ritsumeikan University, Shiga

Motonari UESUGIInstitute for Integrated Cell-Material Sciences/
Institute for Chemical Research, Kyoto University


Educational Background
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

Scientific Interests

B.S. 1990, Kyoto University
Ph.D. 1995, Kyoto University (advisor: Yukio Sugiura)
Postdoctoral Fellow, 1995-1998, Harvard University(advisor: Gregory L. Verdine)
Assistant Professor, 1998-2005, Baylor College of Medicine
Associate Professor (tenured), 2005-2009, Baylor College of Medicine
Professor, 2005-, Kyoto University

Chemical Biology: Discovery, design, synthesis, and biological use of small organic
molecules with unique biological activity

Recent Papers
1.

“A Small Molecule that Promotes Cardiac Differentiation of Human Pluripotent Stem Cells
under Defined Cytokine- and Xeno-free Conditions” Minami, I., et al. Cell Reports in press,
2012.
2. “Synthesis and Evaluation of Diarylthiazole Derivatives that Inhibit Activation of Sterol
Regulatory Element-binding Proteins” Kamisuki, S., et al. J. Med. Chem. 2011, 54(13),4923-4927.
3. “A Mitochondrial Surface-specific Fluorescent Probe Activated by Bioconversion” Kawazoe,
Y., Shimogawa, H., Sato, A., Uesugi, M. Angew. Chem. Int. Ed. 2011, 50(24), 5478-81.
4. “Cell-morphology Profiling of a Natural Product Library Identifies Bisebromoamide andMiuraenamide A as Actin-filament Stabilizers” Sumiya, E., et al. ACS Chem. Biol. 2011,
6(5), 425-31.
5. “Deactivation of STAT6 through Serine 707 Phosphorylation by JNK” Shirakawa, T.
Kawazoe, Y., Tsujikawa, T., Jung, D., Sato, S., Uesugi, M. J. Biol. Chem. 2011, 286, 40034010.
6. “Marine Natural Product Aurilide Activates the OPA1-Mediated Apoptosis by Binding toProhibitin” Sato, S., et al. Chem. Biol. 2011, 18 (1), 131-139.
7. “A Small Molecule that Blocks Fat Synthesis by Inhibiting the Activation of SREBP”
Kamisuki, S., et al. Chem. Biol. 2009, 16 (8), 882-892.
8. “A Dumbbell-shaped Small Molecule that Promotes Cell Adhesion and Growth” Yamazoe,
S., Shimogawa, H., Sato, S., Esko, J. D., Uesugi, M. Chem. Biol. 2009, 16 (7), 773-782.
9. “Wrenchnolol Derivative Optimized for Gene Activation in Cells” Jung, D., Shimogawa, H.,
Kwon, Y., Mao, Q., Sato, S., Kamisuki, S., Kigoshi, H., Uesugi, M. J. Am. Chem. Soc. 2009,
131(13), 4774-4782.
10. “Polyproline-rod Approach to Isolating Protein Targets of Bioactive Small Molecules:
Isolation of a New Target of Indomethacin” Sato, S., et al. J. Am. Chem. Soc. 2007, 129(4),873-880.
11. “Small Molecule Transcription Factor Mimic” Kwon, Y., Arndt, H., Mao, Q., Choi, Y.,
Kawazoe, Y., Dervan, P. B., Uesugi, M. J. Am. Chem. Soc. 2004, 126, 15940-15941.
4th RSC－CSJ Joint Symposium 2013
―Chemical Biology Research by Young Investigators―

9L-5
Small Molecule Tools for
Cell Biology and Cell Therapy
Motonari UESUGIInstitute for Integrated Cell-Material Sciences/Institute for Chemical Research,
Kyoto University, Gokasho, Uji, Kyoto 611-0011 Japanuesugi@scl.kyoto-u.ac.jp

In human history, bioactive small molecules have had three primary uses: as
medicines, agrochemicals, and biological tools. Among them, what our laboratory has
done in the past was the discovery and use of biological tools. Our laboratory has been
discovering and designing small organic molecules with unique activities to them as tools
for biological investigation and manipulation.
In addition to tool discovery, our laboratory has recently become interested in
exploring another application of small molecules: small molecule tools for cell therapy.
Although small molecule drugs will continue to be important, cell therapy will be a
powerful approach to curing difficult diseases that small molecule drugs are unable tohandle. However, there are a number of potential problems in bringing cell therapy
technologies to the clinic, including high cost, potential contamination, low stability, andtumorigenesis. Stable, completely defined small molecule tools, which are usuallyamenable to cost-effective mass production, may be able to help the clinical use of celltherapy.
Through screening chemical libraries, we have been discovering unique synthetic
molecules that modulate or detect fundamental characteristics of human cells useful forcell therapy. Some of such molecules may serve as tools for cell engineering or celltherapy as well as basic cell biological research. This presentation provides a quick
overview of our recent research programs with a special emphasis on the discovery andutilization of “adhesamine.” This dumbbell-shaped synthetic molecule enhanced
attachment and growth of cells by binding to heparan sulfate on cell membrane andthereby clustering syndecan. Using this molecule as a lead, we were able to design small
synthetic molecules with fibronectin-like properties, which boost culture, expansion, andtransplantation of clinically useful cells.
Other small-molecule tools we newly discovered may be discussed in thepresentation as well.

10

The 93rd CSJ Annual Meeting
24th March 2013, Ritsumeikan University, Shiga

Stuart J. CONWAY
Chemistry Research Laboratory
University of Oxford
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Educational Background
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Scientific Interests

B.Sc. 1997, the University of Warwick
Ph.D. 1997-2000, the University of Bristol(advisor: Prof. Jeff Watkins FRS and Dr. David Jane)
Postdoctoral Fellow, 2001-2003, the University of Cambridge(advisor: Prof. Andrew Holmes FRS)
Lecturer, 2003-2008, the University of St Andrews
University Lecturer 2008-present, the University of Oxford
Tutorial Fellow 2008-present, St Hugh’s College, Oxford

Chemical Biology: Discovery, design, synthesis, and biological use of small organic
molecules with unique biological activity

Recent Papers
1.
2.
3.
4.
5.

“Wavelength-orthogonal Photolysis of Neurotransmitters in Vitro” StantonHumphreys et al. Chem. Commun. 2012, 48, 657-659.
“3,5-Dimethylisoxazoles Act as AcetylLysine-mimetic Bromodomain Ligands”
Hewings et al., J. Med. Chem. 2011, 54, 6761-6770. (Highlighted in Faculty 1000).
“Development of Inositol-based Antagonists for the D-myo-Inositol 1,4,5Trisphosphate Receptor” Keddie et al., Chem. Commun. 2011, 47, 242-244.
“Mechanism of Ligand-gated Potassium Efflux in Bacterial Pathogens” Roosild etal., Proc. Natl. Acad. Sci. USA 2010, 107, 19784-19789.
“The Synthesis and Biological Action of Novel 4-Position-modified Derivatives of
D-myo-Inositol 1,4,5-Trisphosphate” Bello et al., J. Org. Chem. 2007, 72, 56475659.

4th RSC－CSJ Joint Symposium 2013
―Chemical Biology Research by Young Investigators―

11

L-6

Cracking the Histone Code:Inhibitors of the BromodomainAcetylLysine Interaction
Stuart J. CONWAY
Department of Chemistry, Chemistry Research Laboratory, University of Oxford
Mansfield Road, Oxford, OX1 3TA, UK
stuart.conway@chem.ox.ac.uk

Lysine acetylation is a key dynamic protein post-translational modification (PTM). Histones, the coreproteins around which DNA is wound, are susceptible to such PTMs (Figure 1A), combinations of which areproposed to form a “histone code” that is involved in regulating gene expression (Figure 1B).1 Lysine
acetylation influences recruitment of transcriptional regulators through interaction with bromodomaincontaining proteins (BCPs), which ‘read’ lysine acetylation state (Figure 1C-E). There are 61 bromodomains,found within 46 proteins; these modules are emerging important therapeutic targets and the protein-protein
interactions they mediate are druggable.2,3 Furthermore, the few BCPs that have been investigated in detail playfundamental cellular roles and show association with specific diseases, including inflammation and cancer.1
Fig. 1. (A) Histones are the core proteins around which nuclear
DNA is packaged. Histone H2A = yellow; Histone H2B = red;Histone H3 = blue Histone H4 = green (PDB ID, 1KX5). (B)Histone acetyl-transferase (HAT) enzymes add an acetyl group
to histone lysines and are viewed as “writers”; bromodomains
bind to and “read” lysine acetylation; histone deacetylases(HDACs) act as “erasers” and remove histone lysines. (C)Bromodomains are a KAc-recognition domain that act asreaders of lysine acetylation state. (D) Bromodomains form acharacteristic four-helix bundle comprising helices αZ, αA, αB
and αC. (E) Recognition of the KAc residue occurs via a directhydrogen bond between the acetyl carbonyl oxygen atom andN1168 (in CREBBP). A second interaction occurs between theacetyl carbonyl oxygen atom and the phenol of Y1125 (inCREBBP) via one of the structured water molecules.

The development of small molecules that selectively prevent the interaction of a specific bromodomain withacetyl-lysine (KAc) is essential to allow dissection of the role that bromodomains play in complex multidomain
proteins. These probes will, ultimately, allow us to better understand the cellular roles of BCPs and are essential
for the validation of bromodomains as therapeutic targets.
We have undertaken studies to develop inhibitors of the BET family of BCPs. An initial screen forbromodomain-binding fragments led to the discovery that the solvent NMP, dihydroquinazolinone and 3,5dimethylisoxazoles (3,5-DMIs) can act as KAc mimics.4 Using a structure-based design approach, we
employed the 3,5-DMI moiety as our lead KAc mimic. Optimisation of the lead compound identified 4 as a
BRD4(1) ligand with an IC50 = 380 nM (AlphaScreen), a KD = 373 nM (SPR), a good selectivity profile overother bromodomains and a ligand efficiency of 0.41 (Figure 2). Compound 4 has been evaluated in a numberof cancer cell lines and has an IC50 = 794 nM in MV4;11 acute myeloid leukaemia cells. This molecule will
be a useful tool for the study of the BET bromodomains. The scaffold reported represents an excellent
prospect for the developmentof further bromodomains
inhibitors.
Fig. 2 The optimisation of the lead 3,5dimethylisoxazole to give a BET-selective
probe with IC50 = 380 nM vs BRD4(1).

1.
2.

Arrowsmith, C. H.; Bountra, C.; Fish, P. V.; Lee, K.; Schapira, M. Nat. Rev. Drug Disc. 2012, 11, 384–400.
Hewings, D. S.; Rooney, T. P. C.; Jennings, L. E.; Hay, D. A.; Schofield, C. J.; Brennan, P. E.; Knapp, S.; Conway, S. J. J. Med.
Chem. 2012, 55, 9393–9413.
3. Conway, S. J. ACS Med. Chem. Lett. 2012, 3, 691–694.
4. Hewings, D. S.; Wang, M.; Philpott, M.; Fedorov, O.; Uttarkar, S.; Filippakopoulos, P.; Picaud, S.; Vuppusetty, C.; Marsden,
B.; Knapp, S.; Conway, S. J.; Heightman, T. D. J. Med. Chem. 2011, 54, 6761–6770.

12

The 93rd CSJ Annual Meeting
24th March 2013, Ritsumeikan University, Shiga

Masayuki INOUE
Graduate School of Pharmaceutical Sciences
The University of Tokyo

Educational Background
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Scientific Interests

B.S. 1993, The University of Tokyo
Ph.D. 1998, The University of Tokyo (advisor: Kazuo Tachibana)
Postdoctoral Fellow, 1998-2000, Sloan-Kettering Institute forCancer Research (advisor: Samuel J. Danishefsky)
Assistant Professor, 2000-2003, Tohoku University (advisor: Masahiro Hirama)
Lecturer, 2003-2004, Tohoku University
Associate Professor, 2004-2007, Tohoku University
Professor, 2007-, The University of Tokyo

Natural Product Synthesis and Bioorganic Chemistry: Synthesis, design and study of
biologically important molecules, with particular emphasis on the total synthesis of
structurally complex natural product

Recent Papers
1.
2.
3.
4.
5.
6.
7.
8.
9.

“Design, Synthesis and Functional Analysis of Dansylated Polytheonamide Mimic: An
Artificial Peptide Ion Channel” H. Itoh, S. Matsuoka, M. Kreir, M. Inoue, J. Am. Chem. Soc.
2012, 134, 14011-14018.
“Radical Amination of C(sp3)–H Bonds Using N-Hydroxyphthalimide and Dialkyl
Azodicarboxylate” Y. Amaoka, S. Kamijo, T. Hoshikawa, M. Inoue, J. Org. Chem. 2012, 77,9959-9969.
“Selective Modification of the N-Terminal Structure of Polytheonamide B Significantly
Changes its Cytotoxicity and Activity as an Ion Channel” N. Shinohara, H. Itoh, S. Matsuoka,
M. Inoue, ChemMedChem, 2012, 7, 1770-1773.
“Photochemically-induced Radical Transformation of C(sp3)H Bonds to C(sp3)CN Bonds” S.
Kamijo, T. Hoshikawa, M. Inoue, Org. Lett. 2011, 13, 5928-5931.
“Application of -Alkoxy Bridgehead Radical for Coupling of Oxygenated Carbocycles” D.
Urabe, H. Yamaguchi, M. Inoue, Org. Lett. 2011, 13, 4778-4781.
“Total Synthesis and Bioactivities of Two Proposed Structures of Maresin” K. Sasaki, D.
Urabe, H. Arai, M. Arita, M. Inoue, Chem. Asian J. 2011, 6, 534-543.
“CCl3CN: A Crucial Promoter of mCPBA-mediated Direct Ether Oxidation” S. Kamijo, S.
Matsumura, M. Inoue, Org. Lett. 2010, 12, 4195-4197.
“Total Synthesis of Polytheonamide B, the Largest Non-ribosomal Peptide” M. Inoue, N.
Shinohara, S. Tanabe, T. Takahashi, K. Okura, H. Ito, Y. Mizoguchi, M. Iida, N. Lee, S.
Matsuoka, Nat. Chem. 2010, 2, 280-285.
“Importance of Twisted Side-chain on Potent Toxicity of Antillatoxin: Total Synthesis andBiological Evaluation of Antillatoxin and Analogs” K. Okura, S. Matsuoka, R. Goto, M. Inoue,
Angew. Chem. Int. Ed. 2010, 49, 329-332.

4th RSC－CSJ Joint Symposium 2013
―Chemical Biology Research by Young Investigators―

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L-7
Total Synthesis and Biological Evaluation of
Polytheonamide B
Masayuki INOUE
Graduate School of Pharmaceutical Sciences, The University of Tokyo,
Hongo, Bunkyo-ku, Tokyo 113-0033, Japaninoue@mol.f.u-tokyo.ac.jp
Polytheonamide B, isolated from the marine sponge Theonella swinhoei, is aposttranslationally modified ribosomal peptide of molecular weight 5030 Da, and displays
extraordinary cytotoxicity. Its 48 amino acid residues include a variety of non-proteinogenic Dand L-amino acids, and the chiralities of these amino acids alternate in sequence. These
structural features induce the formation of a stable β6.3-helix, giving rise to a tubular structure ofover 4 nm in length. In the biological setting, this fold is believed to transport cations across thelipid bilayer through a pore, thereby acting as a transmembrane ion channel.
We firstly developed the total synthesis route to this unusual peptide. This achievement togain precise synthetic control of the structure of polytheonamide B provided the first chemicalbasis for systematically correlating its molecular structure with biological functions.
Specifically, structure-activity relationship (SAR) studies of the 21 fully synthetic variants were
performed to pinpoint the proteinogenic and non-proteinogenic building blocks within themolecule that were essential to the channel function and the toxicity. These achievements
demonstrate the potential benefits of the total synthesis endeavor and the importance of efficientconstruction of the complex molecule. The knowledge accumulated through these studies will
also be useful for rational generation of new tailor-made channel peptides and cytotoxic
molecules with desired functions.

14

The 93rd CSJ Annual Meeting
24th March 2013, Ritsumeikan University, Shiga

Rebecca Jane Miriam GOSS
School of Chemistry, University of St Andrews, UKEducational Background
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Scientific Interests

B.Sc. Hons. Dunelm. 1997, University of Durham
Ph.D. 2001, University of Durham(advisor: Professor David O'Hagan, FRSE)
Postdoctoral Research Associate, 2001-2002, University ofCambridge,(advisors: Professor Peter Leadlay, FRS and Professor Jim Staunton, FRS)
Teaching Fellowship, 2002-2003 Department of Chemistry, University of Nottingham
Royal Society Dorothy Hodgkin Fellowship 2003-2007
Lectureship, School of Chemistry, University of Exeter 2003-closed in 2005
Lectureship, School of Chemistry, University of East Anglia, 2005-2010
Senior Lectureship, School of Chemistry, University of East Anglia, 2011
Readership, School of Chemistry, University of East Anglia, 2012
Reader in Organic/Biomolecular Chemistry, School of Chemistry, University of St Andrews,
2012

The Goss group is interested in the biosynthesis of natural products and in how thesebiosynthetic pathways may be harnessed to generate natural products of our own design.
Many natural products are of medicinal importance. We are also interested in determining themolecular mode of action of drug molecules. It is our aim to couple these two interests,
manipulating biosynthetic pathways to expediently access series of otherwise syntheticallyintractable natural product analogues, which can be utilised in structure activity determination.

Recent Papers
1. “Biogenesis of the Unique Nucleoside of the Uridyl Peptide Antibiotics: Pacidamycin” A. E.
Ragab, S. Grüschow, R. J. M. Goss,* J. Am. Chem. Soc. 2011, 133, 15288-91.
2. “Diversity in Natural Product Families is Governed by More than Enzyme Promiscuity
Alone: Establishing Control of the Pacidamycin Portfolio” S. Grüschow, E. Rackham, R. J.
M. Goss,* Chem. Sci. 2011, 2, 2182-2186.
3. Highlighted in Chemistry World: http://www.rsc.org/chemistryworld/News/2011/August/11081101.asp(Highlight in C&EN News 2011, 89(34), August 22.)
4. “RSC
SelectedHighlightof‘Cutting
Edge
Chemistryin2011”
http://www.rsc.org/chemistryworld/News/2011/December/chemistry-articles-most-excitingevents-2011.asp.
5. “Gene Expression Enabling Synthetic Diver sification of Unnatural Products: Chemogenetic
Generation Pacidamycin Analogs” A. Deb Roy, S. Grüschow, N. Cairns, R. J. M. Goss*, J.
Am. Chem. Soc. 2010, 134, 1224-12245. (Highlighted in C.&EN. News, August 23rd,
2010. )
6. “Pacidamycin Biosynthesis: Identification and Heterologous Expression of the First Uridyl
Peptide Antibiotic Gene Cluster” E. J. Rackham, S. Grüschow, A. E. Ragab, S. Dickens, R.
J. M. Goss*, ChemBioChem. 2010, 11, 1700-1709. (Submitted in February 2010 andpublished online in July 2010, ahead of a manuscript confirming our results, contributed on
August 3rd to PNAS by Professor Christopher Walsh (Harvard))
7. “Direct Evidence for the Use of Multiple Antifungals by a Leaf-cutting Ant” J. Barke, R. F.
Seipke, S. Grüschow, M. J. Bibb, R. J. M. Goss, D. W. Yu, M. I. Hutchings* BMC Biol.
2010, 8, 109. (Labelled “Highly Accessed” http://www.biomedcentral.com/1741-7007/8/109;
5400 downloads in the first month).
4th RSC－CSJ Joint Symposium 2013
―Chemical Biology Research by Young Investigators―

15

L-8
Elucidating and Exploiting Biosynthesis
Rebecca GOSS
School of Chemistry, University of St Andrews
Biomolecular Sciences Building, North Haugh, St Andrews KY16 9ST, UK
rjmg@st-andrews.ac.uk
Natural products represent a treasure trove of medicinally relevant compounds: over the past 3
decades over 70% of antimicrobials and over 60% of antitumor agents entering clinical trials havebeen based on natural products.i Generation of natural product analogues is an important area.
A new paradigm in natural product analogue generation, which we have termed Chemogenetics,
will be described. A genome scanning approach to determining the biosynthetic genes responsible forthe construction of a highly unusual natural product will also be discussed :CHEMOGENETICS a new paradigm in natural product generation
The generation of analogues of natural products is key to understanding structure activity
relationships and improving physicochemical properties. Traditional approaches of analogue
generation such as total synthesis and semisynthesis have limitations. We have pioneered a newconcept in which a gene is introduced to an organism and coerced to work in concert with an existing
biosynthetic pathway. This installs a chemical handle that enables selective derivatisation of thenatural product.

Figure 1. Chemogenetics: gene expression enabling synthetic diversification.

IDENTIFICATION OF THE FIRST URIDYL PEPTIDE ANTIBIOTIC BIOSYNTHETIC
CLUSTER: PACIDAMYCIN
The first identification of the pacidamycin biosynthetic cluster and its heterologous expression,using the cutting edge approach of genome scanning will be described.

Figure 2. Pacidamycin biosynthetic cluster.
i

16

Newman, D. J., Cragg, G. M. J. Nat. Prod. 2007, 70, 461-477.
The 93rd CSJ Annual Meeting
24th March 2013, Ritsumeikan University, Shiga

Kazuya KIKUCHI
Graduate School of Engineering, Osaka University
Immunology Frontier Research Center, Osaka University


Educational Background
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Scientific Interests

B.S. 1988, University of Tokyo
Ph.D. 1994, University of Tokyo (advisor: Masaaki Hirobe)
Postdoctoral Fellow, 1994-1995, University of California, San Diego (advisor: Roger Y.
Tsien)
Postdoctoral Fellow, 1995-1996, The Scripps Research Institute (advisor: Donald Hilvert)
Assistant Professor, 1997-2000, University of Tokyo (advisor: Tetsuo Nagano)
Associate Professor, 2000-2005, University of Tokyo (advisor: Tetsuo Nagano)
Professor, 2005-, Osaka University, Graduate School of Engineering
Professor, 2009-, Osaka University, Immunology Frontier Research Center

Chemical Biology: Design, Synthesis and Application of Molecular Imaging Probes A
short list of interests and activities
Fluorescent Probes, Molecular Imaging, MRI Probes, Optical Imaging, in Vivo Imaging

Recent Papers
1. “Development of Fluorogenic Probe with Transesterification Switch for Detection of Histone
Deacetylase Activity” R. Baba, Y. Hori, S. Mizukami, K. Kikuchi, J. Am. Chem. Soc. 2012,
134, 14310-14313.
2. “Development of Protein Labeling Probes with Redesigned Fluorogenic Switch Based on
Intramolecular Association and No-wash Live-cell Imaging” Y. Hori, K. Nakaki, M. Sato, S.
Mizukami, K. Kikuchi, Angew. Chem. Int. Ed. 2012, 51, 5611-5614.
3. “No-wash Protein Labeling with Designed Fluorogenic Probes and Application to Real-time
Pulse-chase Analysis” S. Mizukami, S. Watanabe, Y. Akimoto, K. Kikuchi, J. Am. Chem. Soc.
2012, 134, 1623-1629.
4. “In Vivo Fluorescence Imaging of Bone-resorbing Osteoclasts” T. Kowada, J. Kikuta, A. Kubo,
M. Ishii, H. Maeda, S. Mizukami, K. Kikuchi, J. Am. Chem. Soc. 2011, 133, 17772-17776.
5. “Covalent Protein Labeling with a Lanthanide Complex and Its Application to
Photoluminescence Lifetime-based Multicolor Bioimaging” S. Mizukami, T. Yamamoto, A.
Yoshimura, S. Watanabe, K. Kikuchi, Angew. Chem. Int. Ed. 2011, 50, 8750-8752.
6. “19F MRI Detection of -Galactosidase Activity for Imaging of Gene Expression” S. Mizukami,
H. Matsushita, R. Takikawa, F. Sugihara, M. Shirakawa, K. Kikuchi, Chem. Sci. 2011, 2, 11511155.
7. “Compound Release System Using Caged Antimicrobial Peptide” S. Mizukami, M. Hosoda, T.
Satake, S. Okada, Y. Hori, T. Furuta, K. Kikuchi, J. Am. Chem. Soc. 2010, 132, 9524-9525.
8. “Photoactive Yellow Protein-based Protein Labeling System with Turn-on Fluorescence
Intensity” Y. Hori, H. Ueno, S.Mizukami, K. Kikuchi, J. Am. Chem. Soc. 2009, 131, 1661016611.
9. “Design, Synthesis and Biological Application of Chemical Probes for Bio-imaging” K.
Kikuchi, Chem. Soc. Rev. 2009, 39, 2048-2053.

4th RSC－CSJ Joint Symposium 2013
―Chemical Biology Research by Young Investigators―

17

L-9
Design, Synthesis and Biological Application of in Vivo
Imaging Probes with Tunable Chemical Switches
Kazuya KIKUCHI
Graduate School of Engineering, Osaka University, Immunology Frontier ResearchCenter, Osaka University 2-1 Yamada-oka, Suita 565-0871, Osaka Japankkikuchi@mls.eng.osaka-u.ac.jp
One of the great challenges in the post-genome era is to clarify the biological significance of
intracellular molecules directly in living cells. If we can visualize a molecule in action, it ispossible to acquire biological information, which is unavailable if we deal with cell homogenates.
One possible approach is to design and synthesize chemical probes that can convert biologicalinformation to chemical output.
Fluorescence protein labeling by synthetic probes is a powerful approach to investigate proteinfunction and localization inside living cells. This chemistry-based technique utilizes a pair of aprotein tag and its specific ligands connected to fluorophores. Its potential advantage is thatvarious fluorescent molecules are available as labeling reagents, and the timing of protein labelingis easily controlled. Because of these characteristics, this method is attracting attention as analternative of fluorescent proteins. On the other hand, in this labeling system, there is a problem
that the fluorescence of free probes inside cells prevents the identification of labeled proteins. Thus,
washing procedures are required to remove the free probes from cells. However, if the probes are
not completely washed out, the remaining probes cause the reduction of the signal-to-noise ratio.
As a solution of this problem, we previously developed a fluorogenic probe for labelingphotoactive yellow protein (PYP) tag. PYP is a small protein (14 kDa) derived from purple
bacteria, and binds to the thioester derivatives of cinnamic acid/coumarin throughtransthioesterification with Cys69. Novel fluorogenic probes, which possess 4-hydroxycinnamic
acid or 7-dimethylaminocoumarin as a ligand scaffold, were synthesized. The labeling kinetics
was significantly improved by these probes. Furthermore, no-wash labeling of intracellular
proteins was successfully achieved. The detail of the strategy for probe design, kinetic experiments
and live-cell imaging will be reported.

18

The 93rd CSJ Annual Meeting
24th March 2013, Ritsumeikan University, Shiga

The 150 Years Anniversary of
UK-Japan Academic Interaction2013 marks the 150th anniversary of the arrival at
UK the Choshu Five from Japan. The five young nobles
from the Choshu clan (Hirobumi Ito, Kaoru Inoue, Yōzō
Yamao, Kinsuke Endo, and Masaru Inoue) smuggled to
London and studied at University College London(UCL) in 1863 in order to receive a mind-broadening
education in readiness for the Meiji restoration.
Furthermore, in 1865, 19 students from the Satsuma clan
arrived at UCL. They would form the core of the newJapanese government – the first of the modern Japan.
On their return they became the first Prime Minister Choshu-Five; the five young nobles fromthe Choshu clan
(Hirobumi Ito), the Minister for Foreign Relations(Kaoru Inoue), Secretary of State in the Ministry of Industries (Yōzō Yamao),founder of the Japanese National Mint (KinsukeEndō), founder of Japanese railways (Masaru
Inoue), the first Minister of Education (Arinori
Mori), and the first head of the University ofTokyo (Yoshinari Hatakeyama). These groups
were followed by many other Japanese students insuccessive years, supported by the Edo Shogunate
Government and then the new Meiji Government,
in a sharing of knowledge and ideas that continues
today.
Statue of the young students fromthe Satsuma Clan
(Kagoshima-city, Japan)

In this history,
two chemists had amost significant contribution to the UK-Japan
interaction in the field of chemistry. Professor
Alexander William Williamson was a Professor
of UCL (1855-1887) and President of the London
Chemical Society, which developed to the Royal
Society of Chemistry. Many of the Japanese
students including the Choshu-five and Satsuma19 lived in Professor. Williamson’s residence.
Professor Williamson looked after all aspects ofuniversity life including registration ofProfessor Alexander William
Williamson

University and arrangement of study subjects. Unfortunately, four Japanese
students died during study in UCL. Professor
Williamson made their graves in the Brookwood
Cemetery, Surrey. Within our 150 Years activities,
Prof. and Mrs. Alexander Williamson’s Monument
Committee has constructed Professor Williamson’smonument at the Brookwood Cemetery.
The Logo of the 150 Years Anniversary of

UK-Japan Academic Interaction
Professor Joji Sakurai became a student of UCL(1876-1881) and studied chemistry under Professor. Williamson. He was an
outstanding student. Thus, he was elected as a fellow of the London Chemical
Society (1879). After returning to Japan, he became the first Professor in the
Department of Chemistry at the University ofTokyo at the age of 25. And he contributed to theestablishment of RIKEN and the Japan Society for
the Promotion of Sciences, and served as Presidentof The Chemical Society of Japan (1883-1885). Also,
he was selected as Fellow of the Japan Academy(Nippon Gakushi-in) and University of London.

UCL, with the support of the Japanese
Embassy in London and the British Embassy inTokyo, is organising a number of events, in the
United Kingdom and Japan to celebrate thebenefits of international understanding that arose
as a result of the arrival in London in 1863 of thefirst Japanese students in UK. Although we will becelebrating these historic events, we will also be
looking forward to the future of the benefits ofwider understanding and cooperation between
Monument in Memory ofProfessor WIliamson
international communities and we are hoping toinitiate a fund to encourage wider interactions between Japanese and UK
students in the future.

Professor Shin-ichi Ohnuma
University College London,
Organising Committee member,
The 150 Years Anniversary of UK-Japan Academic Interaction

4th RSC－CSJ Joint Symposium 2013
―Chemical Biology Research by Young
Investigators―



Date:
Venue:



Hosted by
The Chemical Society of Japan
Royal Society of Chemistry

March 24th (SUN) 9:00‐17:40
Room S8 (CO‐LEARNING HOUSE I C306),
Ritsumeikan University, Biwako‐kusatsu
Campus (Shiga, Japan)

Publication： March 2013
Publisher:

The Chemical Society of Japan

1‐5, Kanda‐Surugadai, Chiyoda‐ku, Tokyo 101‐8307, Japanc The Chemical Society of Japan○