1989 年度実績報告書

血糖降下、血圧降下ペプチドの生合成調節機構

研究課題

研究課題/領域番号	60065004
研究機関	東北大学
研究代表者	岡本宏東北大学, 医学部, 教授 (60025632)
研究分担者	山本博東北大学, 医学部, 助教授 (00115198)
キーワード	VIP遺伝子 / cyclic AMP@phorbol ester-responsive element結合因子 / ペプチドC末端αーアミド化酵素 / reg遺伝子 / rig遺伝子 / pancreatic stone protein / pancreatic thread protein / 酸素添加酵素
研究概要	1.(1)ヒトVIP遺伝子が第6染色体q26→27に局在することをin situハイブリダイゼーションで明らかにした。(2)DNA footーprintingおよびgel retardation法の結果からヒトVIP遺伝子の転写開始点から66ー88塩基対上流に結合する核内因子の存在が見出された。この因子がAキナーゼやCキナーゼでリン酸化されVIPの転写が誘導される可能性が考えられた。(3)ペプチドC末端アミド化酵素をラット脳から精製した。精製酵素は分子量36KでCu^<2+>、アスコルビン酸、41K蛋白を要求する酸素添加酵素と考えられた。 2.(1)ヒトreg遺伝子の全構造を決定した。reg遺伝子は全長2921塩基対で6個のエキソンからなり、転写開始点から27塩基対上流にTATA boxを有していた。この遺伝子構造の解析から、reg蛋白、pancreatic stone protein、pancreatic thread proteinはreg遺伝子によりコードされる同一蛋白であることが明らかとなった。(2)ヒトreg遺伝子は第13染色体に、rig遺伝子は第3ー4染色体に局在することがspot blotハイブリダイゼーションにより推定されたが、ヒトゲノムにはこれらの遺伝子の関連配列や偽遺伝子が多数存在することが明らかになってきたので、これらの偽遺伝子の局在を含めて今後さらに検討する必要が生じてきた。(3)reg蛋白、rig蛋白をそれぞれyeast、大腸菌で大量生産させた。rig蛋白は145個の、reg蛋白はシグナルペプチドが切断された144個のアミノ酸から構成されており、その配列はcDNAから予想したものと一致した。reg蛋白についてはSーS架橋部位やαヘリックス・βシートなどの高次構造も明らかにした。

研究成果
(16件)

すべてその他

すべて文献書誌 (16件)

[文献書誌] Inoue C.et al.: "Nicotinamide prolongs survival of primary cultured hepatocytes without involving loss of hepatocyte-specific functions" J.Biol.Chem.264. 4747-4750 (1989)
[文献書誌] Noguchi,M.et al.: "Rat peptidylglycine α-amidating enzyme:the relation between activities at neural and alkaline pH values" Arch.Biochem.Biophys.275. 505-513 (1990)
[文献書誌] Leffert,J.D.et al.: "Rat amylin:cloning and tissue-specific expression in pancreatic islets." Proc.Natl.Acad.Sci.U.S.A.86. 3127-3130 (1989)
[文献書誌] Sugawara,A.et al.: "Nucleotide sequence determination of mouse,chicken and Xenopus laevis rig cDNAs:the rig-encoded protein is extremely conserved during vertebrate evolution" Biochem.Biophys.Res.Commun.166. 1501-1507 (1990)
[文献書誌] Watanabe,T.et al.: "Complete nucleotide sequence of human reg gene and its expression in normal and tumoral tissues.The reg protein,pancreatic stone protein,and pancreatic thread protein are one and the same product of the gene." J.Biol.Chem.265. (1990)
[文献書誌] Tajima,M.et al.: "The reaction product of peptidylglycine α-amidating enzyme is a hydroxyl derivative at α-carbon of the carboxyl terminal glycine." J.Biol.Chem.265. (1990)
[文献書誌] Shiga,K.et al.: "Isolation and characterization of human rig and its pseudogenes:the functional gene has features characteristic of housekeeping genes." Proc.Natl.Acad.Sci.U.S.A.87. (1990)
[文献書誌] Terazono,K.et al.: "Expression of reg protein in rat regenerating islets and its colocalization with insulin in the β-cell secretory granules" Diabetologia. 33. (1990)
[文献書誌] Kato,I.et al.: "Cloning and expression of pituitary α-amidating enzyme cDNA:variant lacking transmembrane domain" Science. (1990)
[文献書誌] Ito,T.et al.: "Isolation and characterization of human and rat reg proteins produced in Saccharomyces cerevisiae" J.Biol.Chem.(1990)
[文献書誌] Hasegawa,S.et al.: "Nucleotide sequence determination of chicken glucagon precursor cDNA:chicken preproglucagon does not contain glucagon-like peptide II." FEBS Lett.(1990)
[文献書誌] Okamoto,H.: "rig and reg:novel genes activated in insulinomas and in regenerating islets.“Diabetes 1988"(Larkins,R.G.et al.,eds)" Excerpta Medica,Amsterdam, 1243 (1989)
[文献書誌] Okamoto,H.: "The molecular basis of experimental diabetes.“Molecular Biology of the Islets of Langerhans"(Okamoto,H.ed.)" Cambridge University Press,Cambridge, 352 (1990)
[文献書誌] Yamamoto,H.,et al.: "The mosaic evolution of the pancreatic polypeptide gene.“Molecular Biology of the Islets of Langerhans"(Okamoto,H.ed.)" Cambridge University Press,Cambridge, 352 (1990)
[文献書誌] Itoh,N.: "The translational control of proinsulin synthesis by glucose.“Molecular Biology of the Islets of Langerhans"(Okamoto,H.ed.)" Cambridge University Press,Cambridge, 352 (1990)
[文献書誌] Yamagami,T.: "The structure and expression of genes of vasoactive intestinal peptide and related peptides.“Molecular Biology of the Islets of Langerhans"(Okamoto,H.ed.)" Cambridge University Press,Cambridge, 352 (1990)

1989 年度 実績報告書

血糖降下、血圧降下ペプチドの生合成調節機構

研究代表者

岡本 宏 東北大学, 医学部, 教授 (60025632)

研究成果

[文献書誌] Inoue C.et al.: "Nicotinamide prolongs survival of primary cultured hepatocytes without involving loss of hepatocyte-specific functions" J.Biol.Chem.264. 4747-4750 (1989)

[文献書誌] Noguchi,M.et al.: "Rat peptidylglycine α-amidating enzyme:the relation between activities at neural and alkaline pH values" Arch.Biochem.Biophys.275. 505-513 (1990)

[文献書誌] Leffert,J.D.et al.: "Rat amylin:cloning and tissue-specific expression in pancreatic islets." Proc.Natl.Acad.Sci.U.S.A.86. 3127-3130 (1989)

[文献書誌] Sugawara,A.et al.: "Nucleotide sequence determination of mouse,chicken and Xenopus laevis rig cDNAs:the rig-encoded protein is extremely conserved during vertebrate evolution" Biochem.Biophys.Res.Commun.166. 1501-1507 (1990)

[文献書誌] Watanabe,T.et al.: "Complete nucleotide sequence of human reg gene and its expression in normal and tumoral tissues.The reg protein,pancreatic stone protein,and pancreatic thread protein are one and the same product of the gene." J.Biol.Chem.265. (1990)

[文献書誌] Tajima,M.et al.: "The reaction product of peptidylglycine α-amidating enzyme is a hydroxyl derivative at α-carbon of the carboxyl terminal glycine." J.Biol.Chem.265. (1990)

[文献書誌] Shiga,K.et al.: "Isolation and characterization of human rig and its pseudogenes:the functional gene has features characteristic of housekeeping genes." Proc.Natl.Acad.Sci.U.S.A.87. (1990)

[文献書誌] Terazono,K.et al.: "Expression of reg protein in rat regenerating islets and its colocalization with insulin in the β-cell secretory granules" Diabetologia. 33. (1990)

[文献書誌] Kato,I.et al.: "Cloning and expression of pituitary α-amidating enzyme cDNA:variant lacking transmembrane domain" Science. (1990)

[文献書誌] Ito,T.et al.: "Isolation and characterization of human and rat reg proteins produced in Saccharomyces cerevisiae" J.Biol.Chem.(1990)

[文献書誌] Hasegawa,S.et al.: "Nucleotide sequence determination of chicken glucagon precursor cDNA:chicken preproglucagon does not contain glucagon-like peptide II." FEBS Lett.(1990)

[文献書誌] Okamoto,H.: "rig and reg:novel genes activated in insulinomas and in regenerating islets.“Diabetes 1988"(Larkins,R.G.et al.,eds)" Excerpta Medica,Amsterdam, 1243 (1989)

[文献書誌] Okamoto,H.: "The molecular basis of experimental diabetes.“Molecular Biology of the Islets of Langerhans"(Okamoto,H.ed.)" Cambridge University Press,Cambridge, 352 (1990)

[文献書誌] Yamamoto,H.,et al.: "The mosaic evolution of the pancreatic polypeptide gene.“Molecular Biology of the Islets of Langerhans"(Okamoto,H.ed.)" Cambridge University Press,Cambridge, 352 (1990)

[文献書誌] Itoh,N.: "The translational control of proinsulin synthesis by glucose.“Molecular Biology of the Islets of Langerhans"(Okamoto,H.ed.)" Cambridge University Press,Cambridge, 352 (1990)

[文献書誌] Yamagami,T.: "The structure and expression of genes of vasoactive intestinal peptide and related peptides.“Molecular Biology of the Islets of Langerhans"(Okamoto,H.ed.)" Cambridge University Press,Cambridge, 352 (1990)

1989 年度実績報告書

岡本宏東北大学, 医学部, 教授 (60025632)