4年ぶりの同窓会

私の履歴・遍歴• 1973〜1975

東大病院、清瀬結核療養所• 1975〜1982

京大医化学ータンパク精製、有機化学

内科臨床医

トリプトファン代謝、プロスタグランディン代謝、有機合成• 1982〜1984（86）カロリンスカ研究所

５­リポキシゲナーゼ、ロイコトリエンA4水解酵素、有機合成• 1984〜1991

東大栄養学助教授

ロイコトリエン酵素cDNAクローニングから、受容体研究• 1991〜2012

東大生化・分子生物（細胞情報）教授

PAF受容体、ロイコトリエン受容体、LPA受容体、cPLA2研究、アシル転位酵素、リピドミクス• 2012〜2022

国立国際医療研究センター研究所

アシル転位酵素、マウス、ヒトサンプル• 2022〜？

微生物化学研究所

Supervisors of research

Observe nature, from yourown experiments or clinicalexperience

Prof. Osamu Hayaishi(Kyoto)

No new technique, nodiscovery

Prof. Bengt Samuelsson(Stockholm)

3アラキドン酸５­リポキシゲナーゼ
Dual activities, Ca requirement, membranetranslocation (800 x g ppt)
5Arachidonic acid

5-HPETE

LTA45

Purification of a mammalian 5-lipoxygenasefrom rat basophilic leukemia cells
Andrew M. Goetze, Liz Fayer, Jennifer Bouska, DirkBornemeier and George W. Carter

Immunosciences Research, D-47K, Abbott Laboratories, North Chicago, IL 60064, U.S.A.

Abstract
5-Lipoxygenase (5-lipox) has been purified to homogeneity from the 20,000 xg
supernatant of sonicated rat basophilic leukemia (RBL-1) cells using a 4-step procedure.
Purification was achieved primarily through the use of anion-exchange HPLC on two
different media. Using the supernatant from 1 × 10 9 cells, approximately 33 μg of theenzyme can be routinely isolated with an estimated net yield of 5–10%. Purified 5-lipox
consists of a single Mr 73,000 band on SDS gels (reduced or unreduced). When thepurified enzyme was incubated with radiolabeled arachidonic acid and products analyzed
by both straight phase and reversed phase HPLC, 5-hydroperoxyeicosate-traenoic acid(5-HPETE) was the only enzymatic product detected.
Prostaglandins 29 ６８９−７０１．１９８５

A proposed mechanism for LTA4 formationby 5-lipoxygenase

78

生化学講座９０年代

Receptors
PAF receptor, Nature 1991, Neuron, 1992
LTB4 receptors (BLT1, 2), Nature 1997,
J. Exp. Med., 2000, J. Exp. Med, 2005;
Nature CB, 2018
Non-edg LPA4 and LPA6, JBC,2003, JBC,2009
JCI Insight, 2018; Human Mol. Genetics, 2022

S. Ishii, K. Yanagida, K. Noguchi
Snowmass, 2005

Z. Honda, I. Miki(Firenze, 1991)

T. Yokomizo group10

Expression cloning of PAF receptor
the 1st example of lipid GPCRs cloningInduction of BLT in HL-60 cells
ーSubtractive PCR by T. Yokomizo
Northern Blot

[3H] LTB4 binding[3H] LTB4 binding (cpm)

500

non-treatedRA 1µM

400
300
200
1000024(Days of culture)

6Nature, 1997; J. Exp. Med. 2000

Shimizu, T. Annu. Rev. Pharm. Toxic. 49, 123-150, 200913

Phenotypes of cPLA2a (-/-) mice
1.
2.
3.
4.
5.
6.
7.
8.
9.

Reduced symptom of bronchial asthma (Nature,1997).
Decreased mortality & symptoms of ARDS (Nature Immunol, 2001).
Milder symptoms in bleomycin-induced fibrosis (Nature Med, 2002)
Reduced mortality due to thromboembolism （J. Exp. Med, 2002,Blood 2009)Marked reduction of collagen-induced arthritis (J. Exp. Med, 2003).
Milder symptoms in inflammatory bone resorption (J. Exp. Med, 2003)
Milder symptoms in allergic encephalomyelitis (J. Exp. Med 2005, PNAS,2010)
Prevention from atherosclerosis (Amer. J. Physiol. 2012)
Impairment of synaptic plasticity (PNAS, 2010)

Most of phenotypes are explained by the deficiency of downstream lipid
mediators

Kume, Uozumi, Kita

• Phospholipase A2 meetingFailure of cPLA2a inhibitor for clinical use• Collaboration with A-company for 12 years• Screening out a potent and selective inhibitor(20 mg per day, po, good PK and PD)• Target, rheumatoid arthritis, bronchial asthma,osteoporosis (all from animal data)• Stop development in 2014, because of adverse
effects at high doses during phase III clinical trial
in US
17

Turning point in 2003

a year of whole human genome was sequenced,
and 10 years before my formal retirement• How arachidonate is located at sn-2 position,and how membrane diversity is made?

• Need development of comprehensivelipidomics techniques(Ono Pharmaceutical and Shimadzu supported
establishment of a metabolome laboratory at U-Tokyo.)18

2002.10.7

⽇経新聞小野薬品と島津製作所に篤く御礼2005~2010

Reunion after March 11 Great East Japan Earthquake (2011.4.7)

Shimizu, T. Annu. Rev. Pharm. Toxic. 49, 123-150, 200924

Amphipathic properties ofglycerophospholipids useful for lipid bilayer

MicelleLiposome

Stryer, 8thed

Essential for cell structure in aqueous environments
25

Fatty acyl diversity and asymmetry ofglycerophospholipids
sn-1, saturated,monoenesn-2, polyunsaturated
Polarhead• How? Biochemical mechanisms• So what? Biological consequence26

Acyltransferase Family(Diacylglycerol acyltransferase 2)(membranebound Oacyltransferase)27

Update of nomenclature proposal
for LPL acyltransferases

Valentine WJ, , Aoki, J.,, and Shindou, H JBC, 2021
28

29

Scientific activity
LPCAT1
PAFR

BLT1
cPLA2

LPA4

LPCAT2-4h-index, 119. from Google Scholar

国立国際医療研究センター（2012~)• 任期付き年俸制• 優れた動物施設、共通機器、• ナショナルバイオバンク、ヒト検体へのアクセス

2022- 微生物化学研究所（五反田、沼津）

微生物化学研究所の特長と未来• 豊富な天然物資源、ライブラリー、中分⼦化合物• 微⽣物培養、精製技術、⼈材豊富• 構造解析（NMR、X線、電顕、各種のMS)が充実• 創薬に繋がる有機合成、また、沼津での動物実験• 抗菌剤P1、抗NTM薬、神経筋接合部強化剤などの進展• 良いアッセイ系を導⼊、ライブラリーの公開• 抗菌剤開発に国や⺠間の協⼒必要• 神経因性疼痛、抗がん剤などNCGMと共同研究

最後に、多くの方に感謝