1989 年度実績報告書

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

研究課題

研究課題/領域番号	01870017
研究種目	試験研究
研究機関	神戸大学
研究代表者	高井義美神戸大学, 医学部, 教授 (60093514)
研究分担者	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)
キーワード	rasp21 / GTP結合蛋白質 / smgp25 / smgp21 / GDI
研究概要	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。

研究成果
(19件)

すべてその他

すべて文献書誌 (19件)

[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] Mizoguchi,A.: "Localization and subcellular distribution of cellular ras gene products in rat brain." Mol.Brain Res.5. 31-44 (1989)
[文献書誌] Kim,S: "Intrasynaptosomal distribution of the ras rho and smg-25A GTP-binding proteins in bovine brain" Mol.Brain Res.6. 167-176 (1989)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] Shoji,I: "Kinetic analysis of the binding of guanine nucleotide to bovine brain smg p25A." Biochem. Biophys.Res.Commun.162. 273-281 (1989)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] Yamamoto,K.: "Small molecular weight GTP-binding proteins and signal transduction." Clinica.Chimica.Acta.185. 347-356 (1989)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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 年度 実績報告書

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

研究代表者

高井 義美 神戸大学, 医学部, 教授 (60093514)

研究成果

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

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

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

[文献書誌] 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)

[文献書誌] 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)

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

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

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

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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 年度実績報告書

高井義美神戸大学, 医学部, 教授 (60093514)