1989 Fiscal Year Annual Research Report

細胞内プロテア-ゼの遺伝子発現の制御と病態

Research Project

Project/Area Number	63304034
Research Institution	Tokyo Metropolitan Organization for Medical Research
Principal Investigator	鈴木紘一東京都臨床医学総合研究所, 遺伝情報研究部, 部長 (80011948)
Co-Investigator(Kenkyū-buntansha)	杉田秀夫国立精神神経センター, 神経研究所, 所長 (80009951) 加藤敬太郎九州大学, 薬学部, 教授 (70037571) 勝沼信彦徳島大学, 酵素科学センター, 教授 (50035375) 市原明徳島大学, 酵素科学センター, 教授 (20035405) 村地孝京都大学, 医学部, 教授 (10089104)
Keywords	プロテア-ゼ / 遺伝子発現 / カルパイン / プロテアソ-ム / アテプシン / 病態
Research Abstract	プロテア-ゼの発現と病態との関係をカルパイン、プロテアソ-ムなど代表的なプロテア-ゼについて調べた。カルパインの発現は遺伝子の5'上流域にある連続した正負のシス因子で制御されていた。細胞をホルボ-ルエステルで刺激すると同じ部位にあるTREに反応し、カルパインの発現は上昇した。同様なカルパインの発現はHTLV-1の感染によっても観察された、病態時のシグナル伝達のカルパインが関係すると予想された。プロテアソ-ムについては幾つかの分子種の一次構造をラットや酵母の酵素で明らかにし、引き続き全サブユニットの構造を決定すべく研究を進めている。一方RNAレベルでプロテアソ-ムの量がガン細胞で著しく増加する場合が判明し、腫瘍とプロテアソ-ムの関係を含め本プロテア-ゼの生理機能、病態との関係を解析している。プロセシングプロテア-ゼとしては、ラットのゴルジ体膜画分から連続したアミノ酸(塩基性)を見分ける酵素を精製し、性質を調べた。酵母の類似酵素KE×2との比較から、本酵素が哺乳類でプロセシングを行う酵素と判断し引続き検討している。リソゾ-ム中のカテプシンBは同じくリソゾ-ムにあるカテプシンDによって不活性型から活性型に変ることを証明した。また、このカテプシンBは細胞外に放出され、骨のマトリクスを分解し血中のカルシウムを上昇させる。これが昇進すると骨の脱灰と骨粗鬆症を引きおこすが、これがカテプシンBの阻害剤の投与によって防止できる可能性が示唆された。アルツハイマ-病時に蓄積するアミロイドタンパク質の生成に関与するプロテア-ゼを合成ペプチド基質を使って検索し、プロリルエンドペプチダ-ゼとプロテアソ-ムがその候補として同定できた。現在、これらのプロテア-ゼがアミロイドタンパク質前駆体に本当に作用し、アミロイドを生じるか否かを検討している。

Research Products

(23 results)

All Other

All Publications (23 results)

[Publications] M.Nakamura: "Frangmentation of an endogenous inhibitor upon complex formation with high and low calcium-requiring forms of calcum-activated neutral protease" Biochemistry. 28. 449-455 (1989)
[Publications] H.Kawasaki: "Identification and characterization of inhibitor sequence in four repeating domains of the endogenous inhibitor of calcium dependent protease" J.Biochemistry. 106. 274-281 (1989)
[Publications] A.Hata: "Tandemly reiterated negative enhancer-like sequence regulates transcription of a human gene for the large subunit of calcium dependent protease" J.Biol.Chem.264. 6404-6411 (1989)
[Publications] K.Ishidoh: "Gene structure of cathepsin H" FEBS Letters.253. 103-107 (1989)
[Publications] T.Fujiwara: "Molecular cloning of cDNA for proteasomes(Multicatalytic protease complex)from rat liver:Primary structure of the largest subunit" Biochemistry. 28. 7332-7340 (1989)
[Publications] K.Tanaka: "Direct evidence for nuclear and cytoplasmic colocalization of proteasomes(multicatalytic protease complex)in liver" J.Cellular Physiology. 139. 34-41 (1989)
[Publications] W.Matthews: "Involvement of the proteasome in various degradative processes in mammalian cells" Proc.Natl.Acad.Sci.,USA. 86. 2597-2601 (1989)
[Publications] T.Murachi: "Intracellular regulatory system involving calpain and calpastatin" Biochemistry International. 18. 263-294 (1989)
[Publications] T.Tsukahara: "RNA degrading activity is tightly associated with the multicatalytic proteinase ingensin" FEBS Letters. 255. 179-183 (1989)
[Publications] S.Ishiura: "Addition of ATP increases the apparent molecular mass of the multicatalytic proteinase,ingensin" FEBS Letters. 257. 123-126 (1989)
[Publications] S.Ishiura: "Putative N-terminal splitting enzyme of amyloid A4 peptride is the multicatalytic proteinase,ingensin,widely distirbuted among mamalin cells" FEBS Letters. 257. 388-392 (1989)
[Publications] S.Ishiura: "Identification of a putative amyloid A4 generating enzyme as a prolyl endopeptidase" FEBS Letters. 260. 131-134 (1989)
[Publications] T.Takahashi: "Purification and characterization of porcine proline-beta-naphthl amidase,a novel enzyme from intestinal mucosa" J.Biol.Chem.264. 11565-11571 (1989)
[Publications] K.Yanagisawa: "Myelin-associated calpain II" J.Neurochem.51. 803-807 (1989)
[Publications] K.Mizuno: "A unique membrane-bound,calcium dependent endopeptidase with specificity toward paired basic residues in rat liverGolgi Fraction" Biochem.Biophys.Res Commun.164. 780-787 (1989)
[Publications] J.Hino: "Isolation and identification of human brain natriuretic peptides in human cardiac atrium" Biochem.Biophys.Res.Commun.
[Publications] M.Maki: "Inhibition of calpain by a synthetic oligopeptide corresponding to an exon of the human calpastatin gene" J.Biol.Chem.264. 18866-18869 (1989)
[Publications] Murachi,T.: "The calpain-calpastatin system in hepatic cells" Calcium Protein Signaling(Hidaka,H.Ed.,Plenum Press). 445-454 (1989)
[Publications] N.Katunuma: "Possible regulatory mechanisms of intracellular protein catabolism through lysosome" Intracellular Proteolysis(Japan Sci.Soc.Press,Tokyo)(Katunuma,N.,Kominami,E.eds). 3-23 (1989)
[Publications] Ishodoh,K.: "Gene structure of rat cathepsin L" FEBS Letters. 252. 103-107 (1989)
[Publications] T.Niwa: "Studies on the reactive site of the cystatin super family using recombinatnt cystatin A mutant.QVVAG is not essential" FEBS Lettrs. 255. 309-314 (1989)
[Publications] Y.Nishimura: "Evidence that aspartic protease is inbolved in the proteolytic processing event of rat procathepsin L in lysosomes" Arch.Biochen.Biophys.271. 400-406 (1989)
[Publications] Y.Nishimura: "Intracellular processing and activation of lysosomal cathepsin L" Intracellular Proteolysis(N.Ktunuma and E.Kominami,eds.,Jpn Sci.Soc.Press,Tokyo). 61-73 (1989)

1989 Fiscal Year Annual Research Report

細胞内プロテア-ゼの遺伝子発現の制御と病態

Principal Investigator

鈴木 紘一 東京都臨床医学総合研究所, 遺伝情報研究部, 部長 (80011948)

Research Products

[Publications] M.Nakamura: "Frangmentation of an endogenous inhibitor upon complex formation with high and low calcium-requiring forms of calcum-activated neutral protease" Biochemistry. 28. 449-455 (1989)

[Publications] H.Kawasaki: "Identification and characterization of inhibitor sequence in four repeating domains of the endogenous inhibitor of calcium dependent protease" J.Biochemistry. 106. 274-281 (1989)

[Publications] A.Hata: "Tandemly reiterated negative enhancer-like sequence regulates transcription of a human gene for the large subunit of calcium dependent protease" J.Biol.Chem.264. 6404-6411 (1989)

[Publications] K.Ishidoh: "Gene structure of cathepsin H" FEBS Letters.253. 103-107 (1989)

[Publications] T.Fujiwara: "Molecular cloning of cDNA for proteasomes(Multicatalytic protease complex)from rat liver:Primary structure of the largest subunit" Biochemistry. 28. 7332-7340 (1989)

[Publications] K.Tanaka: "Direct evidence for nuclear and cytoplasmic colocalization of proteasomes(multicatalytic protease complex)in liver" J.Cellular Physiology. 139. 34-41 (1989)

[Publications] W.Matthews: "Involvement of the proteasome in various degradative processes in mammalian cells" Proc.Natl.Acad.Sci.,USA. 86. 2597-2601 (1989)

[Publications] T.Murachi: "Intracellular regulatory system involving calpain and calpastatin" Biochemistry International. 18. 263-294 (1989)

[Publications] T.Tsukahara: "RNA degrading activity is tightly associated with the multicatalytic proteinase ingensin" FEBS Letters. 255. 179-183 (1989)

[Publications] S.Ishiura: "Addition of ATP increases the apparent molecular mass of the multicatalytic proteinase,ingensin" FEBS Letters. 257. 123-126 (1989)

[Publications] S.Ishiura: "Putative N-terminal splitting enzyme of amyloid A4 peptride is the multicatalytic proteinase,ingensin,widely distirbuted among mamalin cells" FEBS Letters. 257. 388-392 (1989)

[Publications] S.Ishiura: "Identification of a putative amyloid A4 generating enzyme as a prolyl endopeptidase" FEBS Letters. 260. 131-134 (1989)

[Publications] T.Takahashi: "Purification and characterization of porcine proline-beta-naphthl amidase,a novel enzyme from intestinal mucosa" J.Biol.Chem.264. 11565-11571 (1989)

[Publications] K.Yanagisawa: "Myelin-associated calpain II" J.Neurochem.51. 803-807 (1989)

[Publications] K.Mizuno: "A unique membrane-bound,calcium dependent endopeptidase with specificity toward paired basic residues in rat liverGolgi Fraction" Biochem.Biophys.Res Commun.164. 780-787 (1989)

[Publications] J.Hino: "Isolation and identification of human brain natriuretic peptides in human cardiac atrium" Biochem.Biophys.Res.Commun.

[Publications] M.Maki: "Inhibition of calpain by a synthetic oligopeptide corresponding to an exon of the human calpastatin gene" J.Biol.Chem.264. 18866-18869 (1989)

[Publications] Murachi,T.: "The calpain-calpastatin system in hepatic cells" Calcium Protein Signaling(Hidaka,H.Ed.,Plenum Press). 445-454 (1989)

[Publications] N.Katunuma: "Possible regulatory mechanisms of intracellular protein catabolism through lysosome" Intracellular Proteolysis(Japan Sci.Soc.Press,Tokyo)(Katunuma,N.,Kominami,E.eds). 3-23 (1989)

[Publications] Ishodoh,K.: "Gene structure of rat cathepsin L" FEBS Letters. 252. 103-107 (1989)

[Publications] T.Niwa: "Studies on the reactive site of the cystatin super family using recombinatnt cystatin A mutant.QVVAG is not essential" FEBS Lettrs. 255. 309-314 (1989)

[Publications] Y.Nishimura: "Evidence that aspartic protease is inbolved in the proteolytic processing event of rat procathepsin L in lysosomes" Arch.Biochen.Biophys.271. 400-406 (1989)

[Publications] Y.Nishimura: "Intracellular processing and activation of lysosomal cathepsin L" Intracellular Proteolysis(N.Ktunuma and E.Kominami,eds.,Jpn Sci.Soc.Press,Tokyo). 61-73 (1989)

鈴木紘一東京都臨床医学総合研究所, 遺伝情報研究部, 部長 (80011948)