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小胞体経路の蛋白質選別輸送の分子機構

Research Project

Project/Area Number 04259106
Research Category

Grant-in-Aid for Scientific Research on Priority Areas

Allocation TypeSingle-year Grants
Research InstitutionKyushu University

Principal Investigator

大村 恒雄  九州大学, 名誉教授 (80029933)

Co-Investigator(Kenkyū-buntansha) 三原 勝芳  九州大学, 大学院・医学系研究科, 助教授 (40029963)
姫野 勝  九州大学, 薬学部, 教授 (50037602)
中野 明彦  東京大学, 大学院・理学系研究科, 助教授 (90142140)
勝沼 信彦  徳島文理大学, 健康科学研究所, 教授 (50035375)
池原 征夫  福岡大学, 医学部, 教授 (70037612)
田代 裕  関西医科大学, 学長 (40077558)
高野 達哉  帝京大学, 薬学部, 教授 (40124995)
大西 俊一  京都大学, 理学部, 教授 (00025272)
Project Period (FY) 1992 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥175,400,000 (Direct Cost: ¥175,400,000)
Fiscal Year 1994: ¥50,300,000 (Direct Cost: ¥50,300,000)
Fiscal Year 1993: ¥57,600,000 (Direct Cost: ¥57,600,000)
Fiscal Year 1992: ¥67,500,000 (Direct Cost: ¥67,500,000)
Keywords蛋白質選別輸送 / 小胞体 / ゴルジ装置 / 蛋白質膜透過 / 小胞輸送 / リソソーム / 蛋白質輸送 / エキソサイトーシス / エンドサイトーシス
Research Abstract

真核細胞において粗面小胞体の膜結合リボソームで合成される種々の蛋白質が小胞体からゴルジ装置を経て細胞表層やリソソームに移行する輸送経路を研究対象とし、1)新生ペプチドの小胞体認識機構、2)ペプチドの小胞体膜透過機構、小胞体膜への組み込み機構、3)小胞体からゴルジ装体を経て細胞表層に至る蛋白質の小胞輸送機構、4)小胞輸送においてのゴルジ体の機能、5)小胞輸送過程においての蛋白質の修飾機構、6)リソソームへの蛋白質の選別輸送機構、7)エンドサイトーシス、エキソサイトーシスの分子機構、について分担して研究した。主要な研究実績は次の通りである。1)酵母シグナル認識粒子(SRP)のSRP受容体との相互作用とGTPase活性の制御機構を変異株を用い解析した(天谷)。2)N末にシグナルアンカー配列(SA)をもつP450の膜への組み込み機構を検討し、SAに続くプロリンに富む配列の重要性を確かめた(三原)。C末に膜挿入配列をもつアルデヒド脱水素酵素の小胞体への組み込みシグナルを解析した(山本)。3)小胞体-ゴルジ体間輸送に関与するSarlpの機能を解析しGTPの加水分解が輸送小胞のゴルジ体への移行に必須であることを確かめた(中野)。Rab3Aによる小胞輸送制御を解析し、Rab3Aの標的蛋白質Rabphilin-3Aの結合蛋白質としてGTPcyclohydrolase 1を同定した(佐々木)。4)ゴルジ膜の細胞質側に局在する分子量372kDa、170kDa,99kDaの3種類の蛋白質を同定し一次構造を決定した(池原)5)外来抗原のプロセシングをCathepsin Bが行っていることを証明した(勝沼)。6)リソソームの主要膜成分LGP96がエンドソーム-リソソーム間の融合に関与する証拠を得た(姫野)。自食過程へのユビキチン系の関与を免疫電顕法で確かめた(横田)。7)内皮細胞層でのAngiotensinのトランスサイトーシスへの液胞型H^+ ATPaseの関与を確認した(高野)。

Report

(3 results)
  • 1994 Annual Research Report
  • 1993 Annual Research Report
  • 1992 Annual Research Report
  • Research Products

    (53 results)

All Other

All Publications (53 results)

  • [Publications] K.Oda: "Conversion of secretory proteins into membrane proteins by fusing with a glycosyl-phosphatidylinositol anchor signal of alkaline phosphatase." Biochem.J.301. 577-583 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] M.Sohda: "Molecular cloning and sequence analysis of a human 372-kDa protein localized in the Golgi complex." Biochem.Biophys.Res.Commun.205. 1399-1408 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] N.Katunuma: "Immunological significances of invariant chain from the aspect of its structural homology with cystatin family." FEBS-letter. 349. 265-269 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] A.Mizoguchi: "Localization of Rabphilin-3A on the synaptic vesicle." Biochem.Biophys.Res.Commun.202. 1235-1243 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] K.Imazumi: "Identification of a Rabphilin-3A interacting protein as GTP cyclohydorase I in PC12 cells." Biochem.Biophys.Res.Commun.205. 1409-1416 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] M.Kato: "Phosphorylation of Rabphilin-3A by calmodulin-dependent protein kinase II." Biochem.Biophys.Res.Commun.205. 1776-1784 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] A.Miyazaki: "Comparison of kinetic properties between MSS4 and Rab3A FRF GDP/GTP exchange proteins." FEBS Lett.350. 333-336 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Y.Fujita: "GDP/GTP exchange reaction-stimulating activity of Rabphilin-3A for Rab3A small GTP-binding protein." FEBS Lett.353. 67-70 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] C.Mineo: "Angiotensin II binding activity in cultured porcine arterial endotehlial cells." Biochemical Pharmacology. 48. 1993-1995 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] T.Oka: "Inhibition of GTP hydrolysis by Sarlp causes accumulation of vesicles that are a functionalintermediate of the ER-to-Golgi transport in yeast." J.Cell Biol.124. 425-434 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] K.Fujimura: "The Saccharomyces cerevisiae MSI4 gene encodes the yeast counterpart of componentA of Rab geranylgeranyltransferase." J.Biol.Chem.269. 9205-9212 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] A.Nakano: "Mutationalanalysis of the Sarl protein,a small GTPase which is essential for vesicular transport from the endoplasmic reticulum." J.Biochem.116. 243-247 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] S.Nishikawa: "Inhibition of endoplasmic reticulum(ER)-to-Golgi transport induces relocalization of binding protein(BiP) within the ER to form the BiP bodies." Mol.Biol.Cell.5. 1129-1143 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] K.Goto: "Localization of cathepsin B,D,and L in the rat osteoclast by immuno-light and -electron microscopy." Histochemistry. 101. 33-40 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] T.Kiyoshima: "Immunocytochemical localization of cathepsin L in the synovial lining calls of the rat temporomandibular joint." Archs oral Biol.39. 1049-1056 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] K.Murakami: "The transmembrane region of microsomal cytochrome p450 identified as the endoplasmic reticulum retention signal." J.Biochem.116. 164-175 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] N.Hachiya: "MSF,a novel cytoplasmic chaperone which functions in precursor targeting to mitochondria." EMBO J.13. 5146-5154 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] R.Alam: "cDNA cloning and characterization of mitochondrial import stimulation factor(MSF) purified from rat liver cytosol." J.Biochem.116. 416-425 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] R.Masaki: "Microsomal aldehyde dehydrogenase is localized to the endoplasmic reticulum via its carboxyl-terminal 35 amino acid." J.Cell Biol.126. 1407-1420 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] K.Kambe: "Preferential localization of heat shock protein 47 in dilated endoplasmic reticulum of chicken chondrocytes." J.Histochem.Cytochem.42. 833-841 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] T.Michikawa: "Transmembrane topology and sites of N-glycosylation of inositol 1,4,5-trisphosphate receptor." J.Biol.Chem.269. 9184-9189 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] R-M.Hong: "Association of N-ethyleimide-sensitive factor with synaptic vesicles." FEBS Letters. 350. 253-257 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] S.Yokota: "Formation of autophagosomes during degradation of excess peroxisomes induced by di-(2-ethylhexyl)phthalate tretment.III.Fusion of early autophagosomes with lysosomal compartments." Eur.J.Cell Biol.66. 15-24 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Naomi Hachiya: "A mitochondrial import factor purified from rat liver cytosol is an ATP-dependent conformational modulator for precursor proteins." EMBO J.12. 1579-1586 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Tomoko Kuroiwa: "Detection of a novel 9-kDa endoplasmic reticulum membrane protein in mammalian cells by chemical cross-linking with translocating nascent peptides." J.Biochem.114. 541-546 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Okura Shimozawa: "Core glycosylation of cytochrome P-450(arom)." J.Biol.Chem.268. 21399-21402 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Soh Yamazaki: "Importance of the proline-rich region following signal-anchor sequence in the formation of correct conformation of microsomal cytochrome P-450s." J.Biochem.114. 652-657 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] S.Takahashi: "A mutation of furin causes the lack of the precursor-processing activity in human colon carcinoma LoVo cells." Biochem.Biophys.Res.Commun.195. 1019-1026 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Y.Suzuki: "Dietary regulation of rat intestinal angiotensin-converting enzyme and dipeptidyl peptidase IV." Am.J.Physiol.264. 1153-1159 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] T.Ohsita: "Analysis of lysosomal degradation isothiocyanate-labelled proteins by Toyo-pearl HW-40 affinity chromatography." J.Chromatography. 633. 281-286 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Y.Matsunaga: "Participation of cathepsin B in processing of antigen preseintation to MHC class II." FEBS Lett.324. 325-330 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] T.Imanaka: "Immunological detection of F_1-ATPase gamma-subunit in rat liver peroxisomes." Biochem.Biophys.Res.Commun.195. 1027-1034 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] N.Henomatsu: "Inhibition of intracellular transport of newly synthesized prolactin by bafilomycin Al in a pituitary tumor cell line,GH3 cells." Eur.J.Cell Biol.62. 127-139 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] T.Tagaya: "Correlation between phospholipase A_2 activity and intra-Golgi protein transport reconstituted in a cell-free system." FEBS Lett.324. 201-204 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] S.Koh: "Immunoelectron microscopic localization of the HPC-1 antigen in rat cerebellum." J.Neurocytology. 22. 995-1005 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] S.Nishikawa: "Identification of a gene required for membrane protein retention in the early secretory pathway." Proc.Natl.Acad.Sci.USA. 90. 8197-8183 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] S.Kim: "Purification and characterization of flavine-adenine dinucleotide phosphohydrolase from rat liver lysosomal membranes." J.Biochem.114. 126-131 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] S.Yokota: "Formation of autophagosomes during degradation of excess peroxisomes induced by di-(2-ethylhexyl)phthalate treatment II.Immunocytochemical analysis of early and late autophagosomes." Eur.J.Cell Biol.62. 372-383 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] K.Nishiyama: "ATP-dependent degradation of a mutant serine:pyruvate/alanine:glyoxylate aminotransferase in a primary hyperoxaluria type l case." J.Cell Biol.123. 1237-1248 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Masao Sakaguchi: "Functions of Signal and Signal-Anchor Sequences Are Determined by the Balance between the Hydrophobic Segment and the N-terminal Charge." Proc.Natl.Acad.Sci.USA. 89. 16-19 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Masao Sakaguchi: "Mitochondrial Porin Can Be Translocated across Both Endoplasmic Reticulum and Mitochondrial Membrances." J.Biochem.112. 243-248 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Kazuya Murakami: "An Isozyme of Microsomal Carboxyesterases,carboxyesterase Sec,Is Secreted from Rat Liver into the Blood." J.Biochem.113. 61-66 (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] E.Tsuji: "An active-site mutation(Gly^<633>-Arg)of dipeptidyl peptidase IV causes its retention and rapid degradation in the endoplasmic reticulum." Biochemistry. 31. 11921-11927 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] T.Fujiwara: "Selective cell surface expression of dipeptidyl peptidase IV with mutations at the active site sequence." Biochem.Biophys.Res.Commun.185. 776-784 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] T.Seguchi: "Brefeldin A-resistant mutants of human epidermoid carcinoma cell line with structural changes of the Golgi complex." J.Biol.Chem.267. 11626-11630 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] M.Murata: "pH-dependent membrane fusion and vesiculation of phospholipid large unilamellar vesicles induced by amphiphilic anionic and cationic peptide." Biochemistry. 31. 1986-1992 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] M.Ohashi: "A novel fluorescence method to monitor the lysosomal disinteagation of low density lipoprotein." Eur.J.Cell Biol.59. 116-126 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Kagiwada: "In vitro fusion of rabbit liver Golgi membranes with liposomes." J.Biol.Chem.268. 1430-1435 (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] H.Takemoto: "Heavy chain binding protein(Bip/GRP78)and Endoplasmin are exported from the endoplasmic reticulum in rat exocrine pancreatic cells,similar to protein disulfide-isomerase." Arch.Biochem.Biophys.296. 129-136 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] A.Yokota: "Two forms of the low-affinity Fc receptor for IgE differentially mediate endocytosis and phagocytosis:Identification of the critical cytoplasmic domains." Proc.Natl.Acad.Sci.USA. 89. 5030-5034 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Issay Okazaki: "Purification and Characterization of an 85 kDa Sialoglycoprotein in Rat Liver Lysosomal Membranes." J.Biochem.111. 763-769 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Takahiro Kyoden: "Purification and Characterization of Dipeptidyl Peptidase IV in Rat Liver Lysosomal Membranes." J.Biochem.111. 770-777 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Junnji Ezaki: "Purification and Characterization of(Ca^<2+>-Mg^<2+>)-ATPase in Rat Liver Lysosomal Membranes." J.Biochem.112. 33-39 (1992)

    • Related Report
      1992 Annual Research Report

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Published: 1992-04-01   Modified: 2019-02-28  

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