1989 Fiscal Year Annual Research Report

ras p21類似GTP結合蛋白質のcDNAと抗体の有効利用、特に病態診断への応用

Research Project

Project/Area Number	01870017
Research Category	Grant-in-Aid for Developmental Scientific Research
Research Institution	Kobe University
Principal Investigator	高井義美神戸大学, 医学部, 教授 (60093514)
Co-Investigator(Kenkyū-buntansha)	川原康洋神戸大学, 医学部, 助手 (80169755) 佐野公彦神戸大学, 医学部, 助手 (40205993) 溝口明神戸大学, 医学部, 講師 (90181916) 菊池章神戸大学, 医学部, 助手 (10204827) 貝淵弘三神戸大学, 医学部, 講師 (00169377)
Keywords	rasp21 / GTP結合蛋白質 / smgp25 / smgp21 / GDI
Research Abstract	私共はウシ大脳よりrasp21に類似した全く新しいGTP結合蛋白質(G蛋白質)を2種類精製し、smgp25、smgp21と命名している。これらのG蛋白質は細胞内情報伝達系の重要な一員であり、その機能の解析が種々の病態を理解し、治療法を見出すために必須であると考えられる。これまでに私共はsmgp25とsmgp21のcDNAを単離し、その一次構造を決定している。また、smgp25のmRNAが神経組織に大量に発現しており、PC-12細胞やNB-1細胞の神経様細胞への分化に伴い増加することを見出している。本研究において、私共はsmgp25とsmgp21に対する特異抗体を作成することに成功した。この抗体を用いて、これらのG蛋白質の組織内分布、細胞内分布を検討した結果、smgp25は神経細胞のシナプスや内、外分泌細胞のゴルジ体、分泌顆粒に特異的に分布し、本G蛋白質が、神経伝達物質の放出のみならず、エキソノエンドサイト-シスに深く関与していることが判明した。また、smgp25はPC-12細胞やNB-1細胞の神経様細胞への分化に伴い増加するが、GOTO細胞のグリア様細胞への分化の際には増加しないことから、本G蛋白質が神経系の腫瘍のマ-カ-となる可能性が明らかとなった。一方、smgp21は血小板に大量に分布しており、巨核芽球性白血病(CMK)細胞の血小板様への分化に伴い増加することから、本G蛋白質が血球系細胞の分化マ-カ-となる可能性も明らかとなった。一方、本研究の過程において、私共はsmgp25の活性を制御する全く新しいタイプの蛋白質(smgp25GDI)を発見した。すでに、私共はsmgp25GDIのcDNAを単離し、一次構造を決定すると共に、その特異抗体を作成することにも成功している。今後、これらの抗体やcDNAを用いてsmgp25、smgp21、smgp25GDIと腫瘍の悪性度や組織型との関連を解析する予定である。以上本年度は、予想以上の成果を上げることができた。

Research Products

(19 results)

All Other

All Publications (19 results)

[Publications] Ohmori,T.: "Small molecular weight GTP-binding proteins in human platelet membranes-Purification and characterization of a novel GTP-binding protein with a molecular weight of 22,000." J.Biol.Chem.264. 1877-1881 (1989)
[Publications] Kikuchi,A: "Purification and characterization from bovine brain cytosol of two GTPase-activating proteins specific for smg p21,a GTP-binding protein having the same effector domain as c-ras p21s." J.Biol.Chem.264. 9133-9136 (1989)
[Publications] Kawata,M: "Phosphorylation of smg p21,a ras p21-like GTP-binding protein by cyclic AMP-dependent protein kinase in a cell-free system and in response to prostaglanding E_1 in intact human platelets." J.Biol.Chem.264. 15688-15695 (1989)
[Publications] Mizoguchi,A.: "Localization and subcellular distribution of cellular ras gene products in rat brain." Mol.Brain Res.5. 31-44 (1989)
[Publications] Kim,S: "Intrasynaptosomal distribution of the ras rho and smg-25A GTP-binding proteins in bovine brain" Mol.Brain Res.6. 167-176 (1989)
[Publications] Sano,K: " Tissue-specific expression of a novel GTP-binding protein(smg p25A)mRNA and its increase by nerve growth factor and cyclic AMP in rat pheochromocytoma PC-12 cells." Biochem.Biophys.Res.Commun.158. 377-385 (1989)
[Publications] Ueda,T: "GTPase activating proteins for the sme-21 GTP-binding protein having the same effector domain as the ras proteins in human platelets." Biochem.Biophys.Res.Commun.159. 1411-1419 (1989)
[Publications] Shoji,I: "Kinetic analysis of the binding of guanine nucleotide to bovine brain smg p25A." Biochem. Biophys.Res.Commun.162. 273-281 (1989)
[Publications] Mizoguchi,A.: "Tissue distribution of smg p25A,A ras p21-like GTP-binding protein,studied by use of a specific monoclonal antibody." Biochem.Biophys.Res.Commun.162. 1438 (1989)
[Publications] Kuroda,S.: "Kinetic analysis of the binding of guanine nucleotides to bovine brain rhoB p20,a ras p21-like GTP-binding protein." Biochem.Biophys.Res.Commun.163. 674-681 (1989)
[Publications] Kawata,M.: "Identification of a major GTP-binding protein in boving aortic smooth muscle mrmbranes as smg p21,a GTP-binding protein having the same effector domain as ras p21s." Biochem.Biophys.Res.Commun.163. 1418-1427 (1989)
[Publications] Ohga,N.: "Rabbit intestine contains a protein that inhibits the dissociation of GDP from and the subsequent binding of GTP to rhoB p20,a ras p21-like GTP-binding protein." Biochem.Biophys.Res.Commun.163. 1523-1533 (1989)
[Publications] Yamamoto,K.: "Small molecular weight GTP-binding proteins and signal transduction." Clinica.Chimica.Acta.185. 347-356 (1989)
[Publications] Sasaki,T.: "Purification and characterization from boving brain cytosol of a protein that inhibits the dissociation of GDP from and the subsequent binding of GTP to smg p25A,a ras p21-like GTP-binding protein." J.Biol.Chem.265. 2333-2337 (1990)
[Publications] Hoshijima,M.: "Purification and characterization from bovine brain membranes of a GTP-binding protein with a Mr of 21,000 ADP-ribosylated by an ADP-ribosyltransferase contained in botulinm toxin type C1-Identification as the rhoA gene product." Mol.Brain.Res.7. 9-16 (1990)
[Publications] Matsui,Y.: "Molecular cloning of smg p21B and identification of smg p21 purified from boving brain and human platelets as smg p21B." Biochem.Biophys.Res.Commun.166. 1010-1016 (1990)
[Publications] Hata,Y.: "Inhibition of the ras p21 GTPase-activating protein-stimulated GTPase activity of c-Ha-ras p21 by smg p21 having the same putative effector domain asr as p21s." J.Biol.Chem.inpress (1990)
[Publications] Yamamoto,J.: "A GTPase-activating protein(GAP)for rhoB p20,a ras p21-like GTP-binding protein-Partial purification,characterization and subcellular distribution in rat brain." Mol.Brain.Res.(1990)
[Publications] Kim,S: "Tissue and subcellular distributions of smg p21s ras p21-like GTP-binding proteins having the same putative effector domain as ras p21s, partly distinct from those of c-ras p21s." Mol. Cell.Biol.(1990)

1989 Fiscal Year Annual Research Report

ras p21類似GTP結合蛋白質のcDNAと抗体の有効利用、特に病態診断への応用

Principal Investigator

高井 義美 神戸大学, 医学部, 教授 (60093514)

Research Products

[Publications] Ohmori,T.: "Small molecular weight GTP-binding proteins in human platelet membranes-Purification and characterization of a novel GTP-binding protein with a molecular weight of 22,000." J.Biol.Chem.264. 1877-1881 (1989)

[Publications] Kikuchi,A: "Purification and characterization from bovine brain cytosol of two GTPase-activating proteins specific for smg p21,a GTP-binding protein having the same effector domain as c-ras p21s." J.Biol.Chem.264. 9133-9136 (1989)

[Publications] Kawata,M: "Phosphorylation of smg p21,a ras p21-like GTP-binding protein by cyclic AMP-dependent protein kinase in a cell-free system and in response to prostaglanding E_1 in intact human platelets." J.Biol.Chem.264. 15688-15695 (1989)

[Publications] Mizoguchi,A.: "Localization and subcellular distribution of cellular ras gene products in rat brain." Mol.Brain Res.5. 31-44 (1989)

[Publications] Kim,S: "Intrasynaptosomal distribution of the ras rho and smg-25A GTP-binding proteins in bovine brain" Mol.Brain Res.6. 167-176 (1989)

[Publications] Sano,K: " Tissue-specific expression of a novel GTP-binding protein(smg p25A)mRNA and its increase by nerve growth factor and cyclic AMP in rat pheochromocytoma PC-12 cells." Biochem.Biophys.Res.Commun.158. 377-385 (1989)

[Publications] Ueda,T: "GTPase activating proteins for the sme-21 GTP-binding protein having the same effector domain as the ras proteins in human platelets." Biochem.Biophys.Res.Commun.159. 1411-1419 (1989)

[Publications] Shoji,I: "Kinetic analysis of the binding of guanine nucleotide to bovine brain smg p25A." Biochem. Biophys.Res.Commun.162. 273-281 (1989)

[Publications] Mizoguchi,A.: "Tissue distribution of smg p25A,A ras p21-like GTP-binding protein,studied by use of a specific monoclonal antibody." Biochem.Biophys.Res.Commun.162. 1438 (1989)

[Publications] Kuroda,S.: "Kinetic analysis of the binding of guanine nucleotides to bovine brain rhoB p20,a ras p21-like GTP-binding protein." Biochem.Biophys.Res.Commun.163. 674-681 (1989)

[Publications] Kawata,M.: "Identification of a major GTP-binding protein in boving aortic smooth muscle mrmbranes as smg p21,a GTP-binding protein having the same effector domain as ras p21s." Biochem.Biophys.Res.Commun.163. 1418-1427 (1989)

[Publications] Ohga,N.: "Rabbit intestine contains a protein that inhibits the dissociation of GDP from and the subsequent binding of GTP to rhoB p20,a ras p21-like GTP-binding protein." Biochem.Biophys.Res.Commun.163. 1523-1533 (1989)

[Publications] Yamamoto,K.: "Small molecular weight GTP-binding proteins and signal transduction." Clinica.Chimica.Acta.185. 347-356 (1989)

[Publications] Sasaki,T.: "Purification and characterization from boving brain cytosol of a protein that inhibits the dissociation of GDP from and the subsequent binding of GTP to smg p25A,a ras p21-like GTP-binding protein." J.Biol.Chem.265. 2333-2337 (1990)

[Publications] Hoshijima,M.: "Purification and characterization from bovine brain membranes of a GTP-binding protein with a Mr of 21,000 ADP-ribosylated by an ADP-ribosyltransferase contained in botulinm toxin type C1-Identification as the rhoA gene product." Mol.Brain.Res.7. 9-16 (1990)

[Publications] Matsui,Y.: "Molecular cloning of smg p21B and identification of smg p21 purified from boving brain and human platelets as smg p21B." Biochem.Biophys.Res.Commun.166. 1010-1016 (1990)

[Publications] Hata,Y.: "Inhibition of the ras p21 GTPase-activating protein-stimulated GTPase activity of c-Ha-ras p21 by smg p21 having the same putative effector domain asr as p21s." J.Biol.Chem.inpress (1990)

[Publications] Yamamoto,J.: "A GTPase-activating protein(GAP)for rhoB p20,a ras p21-like GTP-binding protein-Partial purification,characterization and subcellular distribution in rat brain." Mol.Brain.Res.(1990)

[Publications] Kim,S: "Tissue and subcellular distributions of smg p21s ras p21-like GTP-binding proteins having the same putative effector domain as ras p21s, partly distinct from those of c-ras p21s." Mol. Cell.Biol.(1990)

高井義美神戸大学, 医学部, 教授 (60093514)