1990 年度実績報告書

細胞内における蛋白質の高次構造形成と局在化

研究課題

研究課題/領域番号	02304063
研究機関	京都大学
研究代表者	伊藤維昭京都大学, ウイルス研究所, 教授 (90027334)
研究分担者	水永武光東京大学, 農学部, 助手 (30011985) 矢原一郎東京都, 臨床医学総合研究所, 部長 (60109957) 大隅隆姫路工業大学, 理学部, 教授 (50111787) 中野明彦東京大学, 理学部, 講師 (90142140) 水島昭二東京大学, 応用微生物研究所, 教授 (50013313)
キーワード	タンパク質折りたたみ / 細胞内輸送 / 局在化 / シャペロン / 熱ショックタンパク質 / タンパク質膜透過 / 小胞体 / 分泌
研究概要	本総合研究の主要目的である,我国におけるこの分野の研究者の交流,情報交換・討論に関しては,平成2年11月30日,12月1日の2日間にわたり班会議を行った.各班員とも新しい研究成果を含む発表を行い,活発な討論により目的を達成しつつあるものと考える.各班員の研究成果の一端は以下の通りである.合成ペプチド,化学修飾チトクロムC等を用いて,生理的条件下での蛋白質変性状態がモルテン・グロビュ-ル状態であることを示した(後藤).熱ショック蛋白質誘導合成の初期反応に熱ショックσ因子の翻訳レベルでの脱抑制が関与することを見出した(由良).ストレス蛋白質HSP90のサブユニット構成およびグルココルチコイド受容体とアクチン繊維の結合における役割を明らかにした(矢原).蛋白質の核膜透過に関する変異株分離のため酵母のEFー2遺伝子のクロ-ニングと毒素耐性変異の作成を行った(河野).SecE,SecY蛋白質の大量生産,純化および膜透過系の再構成に成功した(水島).変異型SecYのin vitro解析,およびペリプラズム蛋白質の構造形成過程に関する変異株の分離を行った(伊藤).酵母SRH1の条件致死変異を作成し,それが蛋白質分泌に関与していることを見い出した(天谷).膜蛋白質のシグナル・アンカ-配列の機能が,末端の電荷とそれに続く配列の疎水性度に依存することを示した(阪口).ミトコンドリア蛋白前駆体のリポソ-ムへの結合と膜電位の関係を調べた(遠藤).ペルオキシソ-ム型チオラ-ゼの局在化シグナルを同定した(大隅).酵母PDI遺伝子をクロ-ニング,塩基配列決定,増殖必須性の証明を行った(水永).小胞体残留信号KDELの受容体を同定した(吉森).HSP47cDNAをクロ-ニングし一次構造を決定した(平芳).酵母ERーゴルジ関輸送をin vitroで再構成しSEC12,SAR1の役割を確証した(中野).酵母のキラ-因子の分泌におけるプロ配列の重要性を示した(徳永).

研究成果

(40件)

すべてその他

すべて文献書誌 (40件)

[文献書誌] Goto,Y.,Takahashi,N.and Fink,A.L.: "Mechanism of acidーinduced folding of proteins." Biochemistry.29. 3480-3488 (1990)
[文献書誌] Goto,Y.and Fink,A.L.: "Phase diagram for acidic conformational states of apomyoglobin." J.Mol.Biol.214. 803-805 (1990)
[文献書誌] Goto,Y.and Aimoto,S.: "Anionー and pHーdependent conformational transition of an amphiphilic polypeptide." J.Mol.Biol.
[文献書誌] Goto,Y.,Okumura,N.and Aimoto,S.: "ATPーinduced conformational transition of denatured proteins." J.Biochem.
[文献書誌] Nagai,H.,Yano,R.,Erickson,J.and Yura,T.: "Transcriptional regulation of the heat shock regulatory gene rpoH in E.coli:Involvment of a novel cataboliteーsensitive promoter." J.Bacteriol.172. 2710-2715 (1990)
[文献書誌] Miyata,Y.and Yahara,I.: "Cytoplasmic 8Sーglucocorticoid receptor binds actin filaments through the HSP90 moiety." J.Biol.Chem.
[文献書誌] Minami,Y.,Kawasaki,H.,Miyata,Y.,Suzuki,K.and Yahara,I.: "Analysis of native forms and their isoform compositions of the mouse 90ーkDa heat shock protein,HSP90." J.Biol.Chem.
[文献書誌] Sato,T.,Sakaguchi,M.,Mihara,K.and Omura,T.: "The aminoーterminal structures that determine topological orientation of cytochrome Pー450 in microsomal membrane." EMBO J.9. 2391-2397 (1990)
[文献書誌] Kuroiwa,T.,Sakaguchi,M.,Mihara,K.and Omura,T.: "Structural requirments for interruption of protein translocation across rough endoplasmic reticulum membrane." J.Biochem.108. 829-834 (1990)
[文献書誌] Sasaki,S.,Matsuyama,S.and Mizushima,S.: "<In>___ー <vitro>___ー kinetic analysis of the role of the positive charge at the aminoーterminal region of signal peptides in translocation of secretory protein across the cytoplasmic membrane in <Escherichia>___ー coli." J.Biol.Chem.265. 4358-4363 (1990)
[文献書誌] Matsuyama,S.,Kimura,E.and Mizushima,S.: "Complementation of two overlapping fragments of SecA,a protein translocation ATPase of <Escherichia>___ー <coli>___ー,allows ATP binding to its aminoーterminal region." J.Biol Chem.265. 8760-8765 (1990)
[文献書誌] Akita,M.,Sasaki,S.,Matsuyama,S.and Mizushima,S.: "SecA interacts with secretory proteins by recognizing the positive charge at the aminoーterminus of the signal peptide in <Escherichia>___ー <coli>___ー." J.Biol.Chem.265. 8164-8169 (1990)
[文献書誌] Tokuda,H.,Kim,Y.J.and Mizushima,S.: "<In>___ー <vitro>___ー protein translocation into inverted membrane vesicles prepared from <Vibrio>___ー <alginolyticus>___ー is stimulated by the electrochemical potential of Na^+ in the presence of <Escherichia>___ー <coli>___ー SecA." FEBS Letters. 264. 10-12 (1990)
[文献書誌] Mizushima,S.and Tokuda,H.: "<In>___ー <vitro>___ー translocation of bacterial secretory proteins and energy requirements." J.Bioenerg.Biomemb.22. 389-399 (1990)
[文献書誌] Matsuyama,S.,Akimaru,J.and Mizushima,S.: "SecEーdependent overproduction of SecY in <Escherichia>___ー <coli>___ー:Evidence for interaction between two components of the secretory machinery." FEBS Letters. 269. 96-100 (1990)
[文献書誌] Shiozuka,K.,Tani,K.,Mizushima,S.and Tokuda,H.: "The proton motive force lowers the level of ATP required for the <in>___ー <vitro>___ー translocation of a secretory protein in <Escherichia>___ー <coli>___ー." J.Biol.Chem.265. 18843-18847 (1990)
[文献書誌] Tani,K.,Tokuda,H.and Mizushima,S.: "Translocation of proOmpA possessing an intramolecular disulfide bridge into membrane vesicles of <Escherichia>___ー <coli>___ー:Effect of membrane energization." J.Biol.Chem.265. 17341-17347 (1990)
[文献書誌] Tokuda,H.,Shiozuka,K.and Mizushima S.: "Reconstitution of translocation activity for secretory proteins from solubilized components of <Escherichia>___ー <coli>___ー." Eur.J.Biochem.192. 583-589 (1990)
[文献書誌] Shinkai,A.,Akita,M.,Matsuyama,S.and Mizushima,S.: "Quantitative renaturation from a guanidineーdenatured state of the SecA dimer,a 200 kDa protein involved in protein secretion in <Escherichia>___ー <coli>___ー." Biochem.Biophys.Res.Commun.172. 1217-1223 (1990)
[文献書誌] Yamane,K.,Akiyama,Y.,Ito,K.and Mizushima,S.: "A positively charged region is a determinant of the orientation of cytoplasmic membrane proteins in <Escherichia>___ー <coli>___ー." J.Biol.Chem.265. 21166-21171 (1990)
[文献書誌] Akiyama,Y.and Ito,K.: "SecY protein,a membrane embedded secretion factor of E.coli,is cleaved by the OmpT protease in vitro." Biochem.Biophys.Res.Commun.167. 711-715 (1990)
[文献書誌] Akiyama,Y.,Inada,T.,Nakamura,Y.and Ito,K.: "SecY,a multispanning membrane protein,contains a potential leader peptidase cleavage site." J.Bacteriol.172. 2888-2893 (1990)
[文献書誌] Baba,T.,Jacq,A.,Brickman,E.,Beckwith,J.,Taura,T.,Ueguchi,C.,Akiyama,Y.,and Ito,K.: "Characterization of coldーsensitive secY mutants of Escherichia coli." J.Bacteriol.172. 7005-7010 (1990)
[文献書誌] Ito,K.: "Structure,function and biogenesis of SecY,an integral membrane protein involved in protein export." J.Bioenerg.Biomembr.22. 75-78 (1990)
[文献書誌] Nakamura,K.,Takamatsu,H.,Akiyama,Y.,Ito,K.,and Yamane,K.: "Complementation of the protein transport defect of the Escherichia coli secY mutant (secY24) by B.subtilis secY homologue." FEBS Lett.273. 75-78 (1990)
[文献書誌] Ueguchi,C.,and Ito,K.: "Escherichia coli sec mutants accumulate a processed immature form of maltoseーbinding protein (MBP),a lateーphase intermediate in MBP export." J.Bacteriol.172. 5643-5649 (1990)
[文献書誌] Osumi,T.and Fujiki,Y.: "Topogenesis of peroxisomal proteins." BioEssays. 12. 217-222 (1990)
[文献書誌] Katakura,Y.,Mizunaga,T.,Miura,T.and Maruyama,Y.: "The presence of protein disulfide isomerase in yeast saccharomyces cerevisiae." Agr.Biol.Chem.54. 1043-1044 (1990)
[文献書誌] Mizunaga,T.,Katakura,Y.,Miura,T.and Matuyama,Y.: "Purification and characterization of yeast protein disulfide isomerase." J.Biochem.108. 846-851 (1990)
[文献書誌] Kimura,H.,Noiva,R.,Mizunaga,T.,Yamauchi,K.,Horiuchi,R.,Cheng,S.Y.and Lennarz,W.J.: "Thyroid hormone binding proteinb contains glycosylation site binding protein activity." Biochem.Biopys.Res.Commun.170. 1319-1324 (1990)
[文献書誌] Tachikawa,H.,Miura,T.,Katakura,Y.and Mizunaga,T.: "PDII gene is essential for yeast cell growth." J.Biochem.
[文献書誌] Yoshimori,T.: "Protein disulfideーisomerase in rat exocrine pancreatic cells is exported from the endoplasmic reticulum despite possessing the retention signal." J.Biol.Chem.265. 15984-15990 (1990)
[文献書誌] Yoshimori,T.: "The identification of a receptor for the endoplasmic reticulum retention signal,“KDEL",by antiーidiotype antibody." Cell Struct.Funct.15. 444-444 (1990)
[文献書誌] Takemoto,H.: "Secretion of the ER luminal proteins in rat exocrine pancreatic cells." Cell Struct.Funct.15. 444-444 (1990)
[文献書誌] Nakai,A.,Satoh,M.,Hirayoshi,K.and Nagata,K.: "Identification of the ATPーbinding heatーinducible protein of Mr=37,000 as glyceraldehydeー3ーphosphate dehydrogenase." Biochem.Biophys.Res.Commun.
[文献書誌] Hosokawa,N.,Hirayoshi,K.,Nakai,A.,et al.: "Flavonoids inhibit the expression of heat shock proteins." Cell Struc.Funct.15. 393-401 (1990)
[文献書誌] Nakai,A.,Hirayama,C.,Ohtsuka,K.,Hirayoshi,K.and Nagata,K.: "A novel ATPーbinding heat inducible protein of Mr=37,000 that is sensitive to tansfomation in BALB/3T3 cells." J.Cell Physiol.143. 577-589 (1990)
[文献書誌] Nakai,A.,Hirayoshi,K.and Nagata,K.: "Transformation of BALB/3T3 cells by simian virus 40 causes a decreased synthesis of a collagenーbinding protein (hsp47)." J.Biol.Chem.265. 992-999 (1990)
[文献書誌] Utani,A.,Tanaka,T.,(中略),Hirayoshi,K.,et al.: "Another mechanism of the defect in typeIII collagen accumulation into the tissues of a patient with EhlersーDanlos syndorome typeIV:Increased intracellular degradation of typeIII collagen." Lab.Invest.63. 181-188 (1990)
[文献書誌] Tsuru,A.,Nakamura,N.,Takayama,E.,Suzuki,Y.,Hirayoshi,K.and Nagata,K.: "Regulation of the expression of vimentin gene during the defferentiation of mouse myeloid leukemia cells." J.Cell.Physiol.110. 1655-1664 (1990)

1990 年度 実績報告書

細胞内における蛋白質の高次構造形成と局在化

研究代表者

伊藤 維昭 京都大学, ウイルス研究所, 教授 (90027334)

研究成果

[文献書誌] Goto,Y.,Takahashi,N.and Fink,A.L.: "Mechanism of acidーinduced folding of proteins." Biochemistry.29. 3480-3488 (1990)

[文献書誌] Goto,Y.and Fink,A.L.: "Phase diagram for acidic conformational states of apomyoglobin." J.Mol.Biol.214. 803-805 (1990)

[文献書誌] Goto,Y.and Aimoto,S.: "Anionー and pHーdependent conformational transition of an amphiphilic polypeptide." J.Mol.Biol.

[文献書誌] Goto,Y.,Okumura,N.and Aimoto,S.: "ATPーinduced conformational transition of denatured proteins." J.Biochem.

[文献書誌] Nagai,H.,Yano,R.,Erickson,J.and Yura,T.: "Transcriptional regulation of the heat shock regulatory gene rpoH in E.coli:Involvment of a novel cataboliteーsensitive promoter." J.Bacteriol.172. 2710-2715 (1990)

[文献書誌] Miyata,Y.and Yahara,I.: "Cytoplasmic 8Sーglucocorticoid receptor binds actin filaments through the HSP90 moiety." J.Biol.Chem.

[文献書誌] Minami,Y.,Kawasaki,H.,Miyata,Y.,Suzuki,K.and Yahara,I.: "Analysis of native forms and their isoform compositions of the mouse 90ーkDa heat shock protein,HSP90." J.Biol.Chem.

[文献書誌] Sato,T.,Sakaguchi,M.,Mihara,K.and Omura,T.: "The aminoーterminal structures that determine topological orientation of cytochrome Pー450 in microsomal membrane." EMBO J.9. 2391-2397 (1990)

[文献書誌] Kuroiwa,T.,Sakaguchi,M.,Mihara,K.and Omura,T.: "Structural requirments for interruption of protein translocation across rough endoplasmic reticulum membrane." J.Biochem.108. 829-834 (1990)

[文献書誌] Matsuyama,S.,Kimura,E.and Mizushima,S.: "Complementation of two overlapping fragments of SecA,a protein translocation ATPase of <Escherichia>___ー <coli>___ー,allows ATP binding to its aminoーterminal region." J.Biol Chem.265. 8760-8765 (1990)

[文献書誌] Akita,M.,Sasaki,S.,Matsuyama,S.and Mizushima,S.: "SecA interacts with secretory proteins by recognizing the positive charge at the aminoーterminus of the signal peptide in <Escherichia>___ー <coli>___ー." J.Biol.Chem.265. 8164-8169 (1990)

[文献書誌] Mizushima,S.and Tokuda,H.: "<In>___ー <vitro>___ー translocation of bacterial secretory proteins and energy requirements." J.Bioenerg.Biomemb.22. 389-399 (1990)

[文献書誌] Matsuyama,S.,Akimaru,J.and Mizushima,S.: "SecEーdependent overproduction of SecY in <Escherichia>___ー <coli>___ー:Evidence for interaction between two components of the secretory machinery." FEBS Letters. 269. 96-100 (1990)

[文献書誌] Shiozuka,K.,Tani,K.,Mizushima,S.and Tokuda,H.: "The proton motive force lowers the level of ATP required for the <in>___ー <vitro>___ー translocation of a secretory protein in <Escherichia>___ー <coli>___ー." J.Biol.Chem.265. 18843-18847 (1990)

[文献書誌] Tani,K.,Tokuda,H.and Mizushima,S.: "Translocation of proOmpA possessing an intramolecular disulfide bridge into membrane vesicles of <Escherichia>___ー <coli>___ー:Effect of membrane energization." J.Biol.Chem.265. 17341-17347 (1990)

[文献書誌] Tokuda,H.,Shiozuka,K.and Mizushima S.: "Reconstitution of translocation activity for secretory proteins from solubilized components of <Escherichia>___ー <coli>___ー." Eur.J.Biochem.192. 583-589 (1990)

[文献書誌] Shinkai,A.,Akita,M.,Matsuyama,S.and Mizushima,S.: "Quantitative renaturation from a guanidineーdenatured state of the SecA dimer,a 200 kDa protein involved in protein secretion in <Escherichia>___ー <coli>___ー." Biochem.Biophys.Res.Commun.172. 1217-1223 (1990)

[文献書誌] Yamane,K.,Akiyama,Y.,Ito,K.and Mizushima,S.: "A positively charged region is a determinant of the orientation of cytoplasmic membrane proteins in <Escherichia>___ー <coli>___ー." J.Biol.Chem.265. 21166-21171 (1990)

[文献書誌] Akiyama,Y.and Ito,K.: "SecY protein,a membrane embedded secretion factor of E.coli,is cleaved by the OmpT protease in vitro." Biochem.Biophys.Res.Commun.167. 711-715 (1990)

[文献書誌] Akiyama,Y.,Inada,T.,Nakamura,Y.and Ito,K.: "SecY,a multispanning membrane protein,contains a potential leader peptidase cleavage site." J.Bacteriol.172. 2888-2893 (1990)

[文献書誌] Baba,T.,Jacq,A.,Brickman,E.,Beckwith,J.,Taura,T.,Ueguchi,C.,Akiyama,Y.,and Ito,K.: "Characterization of coldーsensitive secY mutants of Escherichia coli." J.Bacteriol.172. 7005-7010 (1990)

[文献書誌] Ito,K.: "Structure,function and biogenesis of SecY,an integral membrane protein involved in protein export." J.Bioenerg.Biomembr.22. 75-78 (1990)

[文献書誌] Nakamura,K.,Takamatsu,H.,Akiyama,Y.,Ito,K.,and Yamane,K.: "Complementation of the protein transport defect of the Escherichia coli secY mutant (secY24) by B.subtilis secY homologue." FEBS Lett.273. 75-78 (1990)

[文献書誌] Ueguchi,C.,and Ito,K.: "Escherichia coli sec mutants accumulate a processed immature form of maltoseーbinding protein (MBP),a lateーphase intermediate in MBP export." J.Bacteriol.172. 5643-5649 (1990)

[文献書誌] Osumi,T.and Fujiki,Y.: "Topogenesis of peroxisomal proteins." BioEssays. 12. 217-222 (1990)

[文献書誌] Katakura,Y.,Mizunaga,T.,Miura,T.and Maruyama,Y.: "The presence of protein disulfide isomerase in yeast saccharomyces cerevisiae." Agr.Biol.Chem.54. 1043-1044 (1990)

[文献書誌] Mizunaga,T.,Katakura,Y.,Miura,T.and Matuyama,Y.: "Purification and characterization of yeast protein disulfide isomerase." J.Biochem.108. 846-851 (1990)

[文献書誌] Kimura,H.,Noiva,R.,Mizunaga,T.,Yamauchi,K.,Horiuchi,R.,Cheng,S.Y.and Lennarz,W.J.: "Thyroid hormone binding proteinb contains glycosylation site binding protein activity." Biochem.Biopys.Res.Commun.170. 1319-1324 (1990)

[文献書誌] Tachikawa,H.,Miura,T.,Katakura,Y.and Mizunaga,T.: "PDII gene is essential for yeast cell growth." J.Biochem.

[文献書誌] Yoshimori,T.: "Protein disulfideーisomerase in rat exocrine pancreatic cells is exported from the endoplasmic reticulum despite possessing the retention signal." J.Biol.Chem.265. 15984-15990 (1990)

[文献書誌] Yoshimori,T.: "The identification of a receptor for the endoplasmic reticulum retention signal,“KDEL",by antiーidiotype antibody." Cell Struct.Funct.15. 444-444 (1990)

[文献書誌] Takemoto,H.: "Secretion of the ER luminal proteins in rat exocrine pancreatic cells." Cell Struct.Funct.15. 444-444 (1990)

[文献書誌] Nakai,A.,Satoh,M.,Hirayoshi,K.and Nagata,K.: "Identification of the ATPーbinding heatーinducible protein of Mr=37,000 as glyceraldehydeー3ーphosphate dehydrogenase." Biochem.Biophys.Res.Commun.

[文献書誌] Hosokawa,N.,Hirayoshi,K.,Nakai,A.,et al.: "Flavonoids inhibit the expression of heat shock proteins." Cell Struc.Funct.15. 393-401 (1990)

[文献書誌] Nakai,A.,Hirayama,C.,Ohtsuka,K.,Hirayoshi,K.and Nagata,K.: "A novel ATPーbinding heat inducible protein of Mr=37,000 that is sensitive to tansfomation in BALB/3T3 cells." J.Cell Physiol.143. 577-589 (1990)

[文献書誌] Nakai,A.,Hirayoshi,K.and Nagata,K.: "Transformation of BALB/3T3 cells by simian virus 40 causes a decreased synthesis of a collagenーbinding protein (hsp47)." J.Biol.Chem.265. 992-999 (1990)

[文献書誌] Utani,A.,Tanaka,T.,(中略),Hirayoshi,K.,et al.: "Another mechanism of the defect in typeIII collagen accumulation into the tissues of a patient with EhlersーDanlos syndorome typeIV:Increased intracellular degradation of typeIII collagen." Lab.Invest.63. 181-188 (1990)

[文献書誌] Tsuru,A.,Nakamura,N.,Takayama,E.,Suzuki,Y.,Hirayoshi,K.and Nagata,K.: "Regulation of the expression of vimentin gene during the defferentiation of mouse myeloid leukemia cells." J.Cell.Physiol.110. 1655-1664 (1990)

1990 年度実績報告書

伊藤維昭京都大学, ウイルス研究所, 教授 (90027334)

[文献書誌] Matsuyama,S.,Kimura,E.and Mizushima,S.: "Complementation of two overlapping fragments of SecA,a protein translocation ATPase of <Escherichia>_ー <coli>_ー,allows ATP binding to its aminoーterminal region." J.Biol Chem.265. 8760-8765 (1990)

[文献書誌] Akita,M.,Sasaki,S.,Matsuyama,S.and Mizushima,S.: "SecA interacts with secretory proteins by recognizing the positive charge at the aminoーterminus of the signal peptide in <Escherichia>_ー <coli>_ー." J.Biol.Chem.265. 8164-8169 (1990)

[文献書誌] Mizushima,S.and Tokuda,H.: "<In>_ー <vitro>_ー translocation of bacterial secretory proteins and energy requirements." J.Bioenerg.Biomemb.22. 389-399 (1990)

[文献書誌] Matsuyama,S.,Akimaru,J.and Mizushima,S.: "SecEーdependent overproduction of SecY in <Escherichia>_ー <coli>_ー:Evidence for interaction between two components of the secretory machinery." FEBS Letters. 269. 96-100 (1990)

[文献書誌] Shiozuka,K.,Tani,K.,Mizushima,S.and Tokuda,H.: "The proton motive force lowers the level of ATP required for the <in>_ー <vitro>_ー translocation of a secretory protein in <Escherichia>_ー <coli>_ー." J.Biol.Chem.265. 18843-18847 (1990)

[文献書誌] Tani,K.,Tokuda,H.and Mizushima,S.: "Translocation of proOmpA possessing an intramolecular disulfide bridge into membrane vesicles of <Escherichia>_ー <coli>_ー:Effect of membrane energization." J.Biol.Chem.265. 17341-17347 (1990)

[文献書誌] Tokuda,H.,Shiozuka,K.and Mizushima S.: "Reconstitution of translocation activity for secretory proteins from solubilized components of <Escherichia>_ー <coli>_ー." Eur.J.Biochem.192. 583-589 (1990)

[文献書誌] Shinkai,A.,Akita,M.,Matsuyama,S.and Mizushima,S.: "Quantitative renaturation from a guanidineーdenatured state of the SecA dimer,a 200 kDa protein involved in protein secretion in <Escherichia>_ー <coli>_ー." Biochem.Biophys.Res.Commun.172. 1217-1223 (1990)

[文献書誌] Yamane,K.,Akiyama,Y.,Ito,K.and Mizushima,S.: "A positively charged region is a determinant of the orientation of cytoplasmic membrane proteins in <Escherichia>_ー <coli>_ー." J.Biol.Chem.265. 21166-21171 (1990)