教員紹介

小笹 徹
Kozasa Tohru

所属・肩書

健康薬学科/生化学研究室、創薬研究センター(教授)

最終学歴

東京大学大学院医学系研究科博士過程修了

取得学位

医学博士

所属学会

日本生化学会、American Society for Pharmacology and Experimental Therapeutics

現在の専門分野

薬理学、生化学

現在の研究課題

●Gタンパク質を介した細胞情報伝達の分子機構

主要研究業績

●Gタンパク質αサブユニットのcDNA及び染色体遺伝子のクローニング
●G12/13-RhoGEF シグナル伝達機構の同定
●Gタンパク質サブユニットの簡便な精製法の開発
●Galphaq-GRK2およびGalphaq-PLCbetaの複合体のX線結晶解析構造の決定
●Galpha13とそのエフェクターであるp115RhoGEFとの複合体のX線結晶解析構造の決定。
●G13伝達系の大腸癌細胞の転移、浸潤での役割の解析

最近の研究業績

1.Patel M, Kawano T, Suzuki N, Hamakubo T, Karginov AV, Kozasa T Galpha13/PDZ-RhoGEF/RhoA signaling is essential for gastrin-releasing peptide receptor-mediated colon cancer cell migration. Mol Pharmacol.: 86: 252-262 (2014)

2.Rusu L, Andreeva A, Visintine DJ, Kim K, Vogel SM, Stojanovic-Terpo A, Chernaya O, Liu G, Bakhshi FR, Haberichter SL, Iwanari H, Kusano-Arai O, Suzuki N, Hamakubo T, Kozasa T, Cho J, Du X, Minshall RD. G protein-dependent basal and evoked endothelial cell vWF secretion. Blood. 2014 123 :442-50 (2014)

3.Gan X, Wang C, Patel M, Kreutz B, Zhou M, Kozasa T, Wu D.Different Raf protein kinases mediate different signaling pathways to stimulate E3 ligase RFFL gene expression in cell migration regulation. J Biol Chem. 288:33978-842013 (2013)

4.Mahajan R, Ha J, Zhang M, Kawano T, Kozasa T, Logothetis DE. A computational model predicts that Gbetagmma acts at a cleft between channel subunits to activate GIRK1 channels. Sci Signal. 6:ra69. (2013)

5.Chow CR, Suzuki N, Kawamura T, Hamakubo T, Kozasa T Modification of p115RhoGEF Ser330 regulates its RhoGEF activity. Cellular Signaling 25:2085-92 (2013)

6.Nance MR, Kreutz B, Tesmer VM, Sterne-Marr R, Kozasa T, Tesmer JJ Structural and Functional Analysis of the Regulator of G Protein Signaling 2-Gα(q) Complex. Structure. 21:438-48. (2013)

7.Gan X, Wang J, Wang C, Sommer E, Kozasa T, Srinivasula S, Alessi D, Offermanns S, Simon MI, Wu D. PRR5L degradation promotes mTORC2-mediated PKC-delta phosphorylation and cell migration downstream of Galpha12. Nat Cell Biol. 14: 686-96 (2012)

8.Hajicek N, Kukimoto-Niino M, Mishima-Tsumagari C, Chow CR, Shirouzu M, Terada T, Patel M, Yokoyama S, and Kozasa T. Identification of critical residues in Galpha13 for stimulation of p115RhoGEF activitiy and the structure of Galpha13-p115RhoGEF RH domain complex. J Biol Chem 286:20625-36 (2011)

9.Chow CR, Douglas Y, Kozasa, T. p115RhoGEF molecule page UCSD-Nature Signaling Gate Way (2011)

10.Waldo GL, Ricks TK, Hicks SN, Cheever ML, Kawano T, Tsuboi K, Wang X, Montel C, Kozasa T, Sondek J, Harden TK. Kinetic scaffolding mediated by a phospholipase C-beta and Gq signaling complex. Science. 330(6006):974-80 (2010)

11.Jernigan KK, Cselenyi CS, Thorne CA, Hanson AJ, Tahinci E, Hajicek N, Oldham WM Lee LA, Hamm HE, Hepler JR, Kozasa T, Linder ME, Lee E. G betagamma activates GSK3 to promote LRP6-mediated beta-catenin transcriptional activity. Sci Signal. 3(121):ra 37 (2010)

12.Gong H, Shen B, Flevaris P, Chow C, Lam S C-T, Kozasa T, Du X. G protein subunit Galpha13 binds to integrin alphaIIb beta3 and mediates integrin “outside-in” signaling. Science 237: 340-343 (2010)

13.Suzuki N, Tsumoto K, Hajicek N, Daigo K, Tokita R, Minami S, Kodama T, Hamakubo T, Kozasa T. Activation of Leukemia-associated RhoGEF by G{alpha}13 with Significant Conformational Rearrangements in the Interface. J Biol Chem. 284:5000-9 (2009)

14.Zhang Y, Chen YH, Bangaru SD, He L, Abele K, Tanabe S, Kozasa T, Yang J.J Origin of the voltage dependence of G-protein regulation of P/Q-type Ca2+ channels. J. Neurosci. 14176-88 (2008)

15.Nicole H, Kozasa, T. Galpha13 molecule page UCSD-Nature Signaling Gate Way A000029 (2008)

16.Shankaranarayanan A. Thal DM, Tesmer VM, Roman DL, Neubig RR, Kozasa T, Tesmer JJ. Assembly of high order Galpha q-effector complexes with RGS proteins. J Biol Chem. 283(50): 34923-34 (2008)

17.Masuho I, Mototani Y, Sahara Y, Asami J, Nakamura S, Kozasa T*, Inoue T. Dynamic expression patterns of G protein-regulated inducer of neurite outgrowth 1 (GRIN1) and its colocalization with Galpha o implicate significant roles of Galpha o -GRIN1 signaling in nervous system. Dev Dyn. ; 237(9):2415-29. (2008) *Corresponding Author

18.Rojas RJ, Yohe ME, Gershburg S, Kawano T, Kozasa T, Sondek J. Galpha-q directly activates p63RhoGEF and Trio via a conserved extension of the DH-associated PH domain. J Biol Chem. 282:29201-10. (2007)

19.Kreutz B, Hajicek N, Yau DM, Nakamura S, Kozasa T. Distinct regions of Galpha13 participate in its regulatory interactions with RGS homology domain-containing RhoGEFs. Cell Signal. 8, 1681-9 (2007).

20.Kawano T, Zhao P, Floreani C V, Nakajima Y, Kozasa T*, Nakajima S*. Interaction of Galphaq and Kir3 (GIRK) Channels. Mol Pharm. 71, 1179-84. (2007) * co-corresponding author

21.Huang JS, Dong L, Kozasa T, Le Breton GC. Signaling through G(alpha)13 switch region I is essential for protease-activated receptor 1-mediated human platelet shape change, aggregation, and secretion. J Biol Chem. 282, 10210-22 (2007).

22.Zhao Q, Albsoul-Younes AM, Zhao P, Kozasa T, Nakajima Y, Nakajima S. Dominant negative effects of a Gbeta mutant on G-protein coupled inward rectifier K+ channel. FEBS Lett. 58, 3879-82 (2006)

23.Marty C, Kozasa T, Quinn MT, Ye RD. Activation state-dependent interaction between Galpha i and p67phox. Mol Cell Biol. 26, 5190-200 (2006).

24.Kreutz B, Yau DM, Nance MR, Tanabe S, Tesmer JJ, Kozasa T. A New Approach to Producing Functional Galpha Subunits Yields the Activated and Deactivated Structures of Galpha(12/13) Proteins. Biochemistry 45,167-174 (2006).

25.Tesmer VM, Kawano T, Shankaranarayanan A, Kozasa T, Tesmer JJ. Snapshot of activated G proteins at the membrane: the Galphaq-GRK2-Gbetagamma complex. Science 310, 1686-90 (2005).

26.Clancy SM, Fowler CE, Finley M, Suen KF, Arrabit C, Berton F, Kozasa T, Casey PJ, Slesinger PA. Pertussis-toxin-sensitive Galpha subunits selectively bind to C-terminal domain of neuronal GIRK channels: evidence for a heterotrimeric G-protein-channel complex. Mol Cell Neurosci. 28, 375-89 (2005).

27.Kino T, Kozasa T, Chrousos GP. Statin-induced blockade of prenylation alters nucleocytoplasmic shuttling of GTP-binding proteins gamma2 and beta2 and enhances their suppressive effect on glucocorticoid receptor transcriptional activity. Eur J Clin Invest. 35, 35508-13 (2005).

28.Nakata H, Kozasa T. Functional characterization of Galphao signaling through G protein-regulated inducer of neurite outgrowth 1. Mol Pharmacol. 67, 695-702 (2005)

29.Nakamura, S., Kreutz, B., Tanabe, S., Suzuki, N., Kozasa, T. Critical role of lysine 204 of Galpha13 to regulate p115RhoGEF and LARG. Mol. Pharmacol 66,1029-1034 (2004).

30.Wang, Q., Liu, M., Kozasa, T., Rothstein, J.D., Sternweis, P.C., and Neubig, R.R. Thrombin and LPA receptors utilize distinct RhoGEFs in Prostate Cancer Cells. J. Biol. Chem. 279, 28831-4 (2004).

31.Yau, D.M., Yokoyama, N., Goshima, Y., Siddiqui, Z.K., Siddiqui, S.S., and Kozasa, T. Identification amd molecular characterization of Galpha12-RhoGEF pathway in C. elegans. Proc. Natl. Acad. Sci. USA 100, 14748-14753 (2003)

32.Johnson, E.N., Seasholtz, T.M., Kreutz, B., Suzuki, N., Kozasa, T., Brown, J.H., and Druey, K.M. RGS16 inhibits signaling through the Galpha13-Rho axis. Nature Cell Biol. 12, 1095-103 (2003).

33.Zhao, Q., Kawano, T., Nakata, H., Nakajima, Y., Nakajima, S., and Kozasa, T. Interaction of G protein beta subunit with inward rectifier K+ Channel Kir3. Mol. Pharmacol. 64, 1085-1091 (2003).

34.Suzuki, N., Nakamura, S., Mano, H., and Kozasa, T. Galpha12 activates RhoGTPase through tyrosine phosphorylated leukemia-associated RhoGEF (LARG). Proc. Natl. Acad. Sci. U.S.A. 100, 733-738 (2003)

35.Manganello, J., Huang, J-S., Kozasa, T., Voyno-Yasenetskaya, T. and Le Breton, G.C. Switch region I of Galpha13 is phosphorylated by cAMP-dependent kinase. J. Biol. Chem. 278, 124-30 (2003)

36.Wells, D.C., Liu, M-Y, Jackson, M., Gutowski, S., Sternweis, P.M., Rothstein, J.D., Kozasa, T., and Sternweis, P. C. Mechanisms for reversible regulation between p115RhoGEF and GTRAP48 and the G12 class of heterotrimeric G proteins. J. Biol. Chem. 277, 1174-81 (2002)

37.Rochdi, M.D., Watier, V., La Madeleine, C., Nakata, H., Kozasa, T., Parent, J.L. Regulation of GTP-binding protein alpha q (Galpha q) signaling by the ezrin-radixin moesin-binding phosphoprotein-50 (EBP50). J. Biol. Chem. 277, 40751-9 (2002).

38.Albsoul-Younes, A.M., Sternweis, P.M., Zhao, P., Nakata, H., Nakajima, S., Nakajima, Y., and Kozasa, T. Interaction sites of the G protein beta subunit with brain GIRK. J. Biol. Chem. 276, 12712-12717 (2001)

39.Niu, J., Vaiskunaite, R., Suzuki, N., Kozasa, T., Carr, D.W., Dulin, N., Voyno Yasenetskaya, T.A. A-kinase anchoring protein 110 (AKAP110): A novel effector molecule of G13 protein that mediates cAMP-independent PKA activation. Current Biology 11, 1686-90 (2001)

40.Shepard, L.W., Yang, M., Xie, P., Browning, D.D., Voyno-Yasenetskaya, T., Kozasa, T., and Ye, R.D. Constitutive activation of NF-{kappa}B and secretion of IL-8 induced by the G protein-coupled receptor of Kaposi's sarcoma-associated herpesvirus involves G{alpha}(13) and RhoA. J Biol Chem. 276, 45979-87 (2001)

41.Vaiskunaite, R., Kozasa, T., and Voyno-Yasenetskaya T. A. Interaction between Galpha subunit of heterotrimeric G12 protein and heat shock protein90, Hsp90 is required for G12 signaling. J. Biol. Chem. 276, 46088-93 (2001)

42.Moratz, C., Kang, V. H., Druey, K. M., Shi, C. S., Scheschonka, A., Murphy, P. M. Kozasa, T., and Kehrl, J. H. Regulator of G Protein Signaling 1 (RGS1) Markedly Impairs Gi Signaling Responses of B Lymphocytes. J. Immunology 164, 1829-1838 (2000)

43.Shi, C.S., Sinnarajah, S., Cho, H., Kozasa, T., Kehrl, J.H. Galpha13 -mediated PYK2 Activation. PYK2 is a mediator of Galpha13 - induced serum response element dependent transcription. J. Biol. Chem. 275, 24470-24476 (2000)

44.Vaiskunaite, R., Adarichev, V., Furthmayr, H., Kozasa, T., Gudkov, A.,Voyno Yasenetskaya, T.A. Conformational activation of radixin by G13 protein {alpha} subunit. J. Biol. Chem. 275, 26206-26212 (2000)

45.Cho, H., Kozasa, T., Takekoshi, K., De Gunzburg, J., Kehrl, J.H. RGS14, a GTPase-activating protein for Galpha i attenuated Galpha i- and Galpha13-mediated signaling pathways. Mol. Pharmacol. 58, 569-576 (2000)

46.Carman, C.V., Parent, J-L., Day, P.W., Pronin, A.N., Sternweis, P.M., Wedegaertner, P.B., Gilman, A.G., Benovic, J.L., and Kozasa, T. Selective Regulation of Galpha q/11 by an RGS domain in the G protein-coupled receptor kinase, GRK2. J. Biol. Chem. 274, 34483-34492 (1999)

47.Chen, L.T., Gilman, A.G., and Kozasa, T. A candidate target for G protein action in brain. J. Biol. Chem. 274, 26931-26938 (1999)

48.Lin, C.H., Duncan, J.A., Kozasa, T., and Gilman A.G. Sequestration of G-protein betagamma subunit inhibits receptor mediated endocytosis. Proc. Natl. Acad. Sci U.S.A. 95, 5057-5060 (1998)

49.Kozasa, T., Jiang, X., Hart, M.J., Sternweis, P.M., Singer, W.D., Gilman, A.G., Bollag, G., Sternweis, P.C. p115 RhoGEF is a GTPase activating protein for Galpha12 and Galpha13. Science 280, 2109-2111 (1998)

50.Hart, M.J., Jiang, X., Kozasa, T., Roscoe, W., Singer, W.D., Gilman, A.G., Sternweis, P.C., Bollag, G. Galpha13 directly stimulates the nucleotide exchange activity of p115 RhoGEF. Science 280, 2112-2114 (1998)

51.Hepler, J.R., Berman, D.M., Gilman, A.G. and Kozasa, T. RGS4 and GAIP are GTPase-activationg proteins for Galphaq and block activation of phospholipase C-beta by gamma-thio-GTP-Galphaq Proc. Natl. Acad. Sci. USA 94, 428-432 (1997)

52.Kozasa, T. and Gilman, A.G. Protein kinase C phosphorylates Galpha12 and inhibits its interaction with Galphaz J. Biol. Chem. 271, 12562-12567 (1996)

53.Berman, D.M., Kozasa, T., and Gilman, A.G. The GTPase-activating protein RGS4 stabilizes the transition state for nucleotide hydrolysis. J. Biol. Chem. 271, 27209-27212 (1996)

54.Nakajima, Y., Nakajima, S., and Kozasa, T. Activation of G protein -coupled inward rectifier K+ channels in brain neurons requieres association of G protein betagannma subunits in cell membranes. FEBS Lett. 390, 217-220 (1996)

55.Kozasa, T. and Gilman, A.G. Purification of recombinant G Proteins from Sf9 Cells by hexa-histidine tagging of associated subunits : characterization of alpha12 and inhibition of adenylyl cyclase by alphaz. J. Biol. Chem. 270, 1734-1741 (1995)

56.Hepler, J. R., Kozasa, T., Smrcka, A. V., Simon, M. I., Rhee, S. G., Sternwise, P. C. and Gilman, A.G. Purification from Sf9 cells and characterization of recombinant Galphaq and Galpha11 . J. Biol. Chem., 268, 14367-14375 (1993)

57.Kozasa, T., Hepler, J.R., Smrcka, A.V., Simon, M. I., Rhee, S. G., Sternwise, P. C., and Gilman, A. G. Purification and characterization of recombinant Galpha16 from Sf9 cells: activation of purified phospholipase C isozymes by G-protein alpha subunits. Proc. Natl. Acad. Sci. USA, 90, 9176-9180 (1993)

58.Kozasa, T., Itoh, H., Tsukamoto, T. and Kaziro, Y. Isolation and characterization of human Gsalpha gene. Proc. Natl. Acad. Sci. USA, 85, 2081-2085 (1988)

59.Itoh, H., Toyama, R., Kozasa, T., Tsukamoto, T., Matsuoka, M. and Kaziro, Y. Presence of three distinct molecular species of Gi protein alpha subunit : structure of rat cDNAs and human genomic DNAs. J. Biol. Chem., 263, 6656-6664 (1988)

60.Matsuoka, M., Itoh, H., Kozasa, T. and Kaziro, Y. Sequence analysis of cDNA and genomic DNA for a putative pertussis toxin-insensitive guanine nucleotide-binding regulatory protein alpha subunit. Proc. Natl. Acad. Sci. USA, 85, 5384-5388 (1988)

61.Fukui, Y., Kozasa, T., Kaziro, Y., Takeda, T.,and Yamamoto, M. Role of a ras homologue in the life cycle of Schizosaccaromyces Pombe. Cell, 44, 329-336 (1986)

62.Itoh, H., Kozasa, T., Nagata, S., Nakamura, S., Katada, T., Ui, M., Iwai, S., Ohtsuka, E., Kawasaki, H., Suzuki, K. and Kaziro, Y. Molecular cloning and sequence determinatoon of cDNAs for alpha subunits of the guanine nucleotide-binding proteins Gs, Gi, and Go from rat brain. Proc. Natl. Acad. Sci. USA, 83, 3776-3780 (1986)

受賞

●Human Frontier Science Program Longterm Fellowship (1991-1993)
●American Heart Association Established Investigator (2001-2005)

著書及び総説

1.小笹 徹  構造から創薬に向かうGPCR研究 実験医学 2013年2月号  企画及び総説

2.Kozasa T, Hajicek N, Chow CR, Suzuki N. Signalling mechanisms of RhoGTPase regulation by the heterotrimeric G proteins G12 and G13. J Biochem. 150:357-69 (2011)

3.Baba M, Kozasa T, Hamakubo T, Kuroda T, Masuda K, Yoneya S, and Kodama T. Soft X-ray Laser Microscopy of Lipid Rafts Towards GPCR-based Drug Discovery Using Time-Resolved FRET Spectroscopy Pharmaceuticals 4:1-26 (2011)

4.Kozasa T Mechanism of RhoGTPase Activation through GPCR : GCPR and Cancer Progression Igakunoayumi (invited review, in Japanese) 233:265-271 (2010)

5.Hajicek N, Kreutz B, Kozasa T Signaling through Galpha12/13 and RhoGEFs. The RhoGTPase in Cancer ed. by van Golen Springer (2010)

6.Suzuki N, Hajicek N, Kozasa T. Regulation and physiological functions of G12/13-mediated signaling pathways. Neurosignals 17: 55-70 (2009).

7.Kozasa, T. Synthetic GEFs for RhoGTPases: A strategy to unwind and assemble the signaling network to control cellular morphology. Cell Science Reviews 4(2) 31-34 (2007)

8.Tanabe, S., Kreutz, B. and Kozasa, T. Biochemical and functional analysis of RGS-RhoGEFs. Methods Enzymol. Regulators of G protein Signaling 390, 285-94 (2004)

9.Iida N, Kozasa T. Identification and biochemical analysis of GRIN1 and GRIN2. Methods Enzymol. Regulators of G protein Signaling 390, 475-83 (2004).

10.Wang, Q., Liu, M., Kozasa, T., Rothestein, J.D., Sternweis, P.C. and Neubig, R.R. Ribozyme- and siRNA-Mediated Suppression of RGS-containing RhoGEF Proteins. Methods Enzymol. 389, 244-65 (2004).

11.Kozasa, T. The structure of GRK2-Gbetagamma complex: Intimate association of G protein signaling modules. Trends Pharmacol Sci. 25, 61-63 (2004)

12.Kozasa, T. and Ye, R. Selective inhibition of G protein-mediated pathways using RGS domain. Methods In Molecular Biology ed. by A. Smrcka Humana Press 237, 153-168 (2004)

13.Kozasa, T. Purification of G protein subunits using Sf9-baculovirus expression system. Methods In Molecular Biology ed. by A. Smrcka Humana Press 237, 21-38 (2004)

14.Kozasa, T. The mechanism of Gbetagamma effector interaction. Handbook of Cell Signaling Vol. 2, Academic Press 639-643 (2003)

15.Kozasa, T. Regulation of G protein-mediated signal transduction by RGS proteins. Life Sci. 68, 2309-2317 (2001)

16.Kozasa, T. Expression and Purification of G protein and alpha subunits from Sf9 cells. “ G proteins :Analysis of Technique” ed. by D. R. Manning CRC 23-38 (1999)

17.Kozasa, T. : Regulation of G protein function by RGS protein (invited review) Seikagaku (in Japanese) 70, 1418-22 (1998)

18.Kaziro, Y., Itoh, H., Kozasa, T., Nakafuku, M., and Satoh, T. Structure and function of signal transducing GTP-binding proteins. Annu. Rev. Biochem. 60, 349-440 (1991)

入試情報 ADMISSION