1998 Fiscal Year Annual Research Report

抗生物質に特異性を有する透過チャンネルの原子構造と機能

Research Project

Project/Area Number	10044320
Research Category	Grant-in-Aid for international Scientific Research
Section	Joint Research .
Research Institution	Tokai University
Principal Investigator	中江太治東海大学, 医学部, 教授 (50102851)
Co-Investigator(Kenkyū-buntansha)	GABRIEL Rumm University of Basel, Biozentrum, Research A TILMAN Schir University of Basel, Biozentrum, Professor JURG P Rosen University of Basel, Biozentrum, Professor 米山裕東海大学, 医学部, 講師 (10220774) 良原栄策東海大学, 医学部, 助教授 (70167063)
Keywords	膜蛋白 / 結晶 / X線回析 / 透過性 / 排出 / 抗生物質 / ポンプ / 耐性
Research Abstract	病院内感染起因菌である緑膿菌は抗生物質に特異性を有するOprD蛋白を発現すると共に抗生物質を細胞外に排出するポンプ蛋白を発現する。本研究ではこれらの蛋白質の機能解析のため原子レベルの構造を明らかとしようとするものである。先ずOprD及びOprM蛋白質を高度に発現する目的で、すでにクローン化されているoprD及びoprM遺伝子を高発現ベクターに構築した。更にOprMに関しては精製を容易にする目的でHis標識を付けた。これらの膜蛋白質はoctyl-β-glucosideで可溶化しイオン交換クロマト及びアフィニティクロマトによって均一にまで精製した。精製標品はバーゼル大学に持ち込み種々の条件下に結晶化を試みたところOprDは50mMMops-500mMLiCl-30%ポリエチレングリコール-0.6%decanoyl sncroseの中で結晶か得られた。OprMは50mMMops-500mMLiCl-15%ポリエチレングリコール-0.6%octyl-2-hydroxyethyl sulfoxide-0.1%octyl polyoxyethylenglycolの中で結晶を得ることか出来た。今後はこれらのX線回析をすべく準備中である。そのため東海大学にX線解析装置を設置した。Schirmer,Tilmanはその始動を手伝うため東海大学に招へいされた。またRunmel,Gabrielは結晶化技術を更に高めるため東海大学に招へいされた。

Research Products
(20 results)

All Other

All Publications (20 results)

[Publications] Nakae,T.: "Subunit Swapping in the Mex-Extrusion Pumps in Pseudomonas aeruginosa" BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS. 244. 898-902 (1998)
[Publications] 中江太治: "緑膿菌の多剤交叉耐性と薬剤排出" 現代医療. 30. 1249-1254 (1998)
[Publications] Taiji NAKAE,: "Flocculation and Membrane Binding of Outer Membrane Protein F,Porin,at Acidic pH" Jpn.J.Appl.Phys. 37. 2074-2077 (1998)
[Publications] Taiji Nakae: "Identification of the Catalytic Triad of the protein D2 Protease in Pseudomonas aeruginosa" BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS. 247. 142-145 (1998)
[Publications] Taiji Nakae: "Antibiotic stress induces a large amount of outer membrane protein in Pseudomonas aeruginosa" FEMS Microbiology Letters. 165. 261-265 (1998)
[Publications] 中江太治: "非特異的抗生物質耐性の機序" Clinical View. 4. 4-5 (1998)
[Publications] Rosenbusch,J.P: "Coupling site-directed mutagenesis with high-level expression:large scale production of mutant porins from Escherichia coli." FEMS Microbiology Letters. 163. 65-72 (1998)
[Publications] Rosenbusch,J.P.: "Lipidic Cubic Phases:New Matrices for the Three-Dimensional Crystallization of Membrane Proteins." J.Struct.Biol.121. 82-91 (1998)
[Publications] Schirmer,T.: "Crystal structure of the IIB subunit of a fructose permease(IIBLev)from Bacillus subtilis." J.Mol.Biol.276. 591-602 (1998)
[Publications] Schirmer,T: "Crystal structure of human recombinant omithine aminotransferase." J.Mol.Biol. 277. 81-102 (1998)
[Publications] Rummel,G.,Schirmer,T: "Detergent binding in trigonal crystals of OmpF porin from Escherichia coli." Biochimie. 80. 543-551 (1998)
[Publications] Schirmer,T: "Stability of trimeric OmpF porin:the contributions of the latching loop L2" Biochemistry. 37. 15663-15670 (1998)
[Publications] Rosenbusch,J.P.: "Overexpression of outer membrane proteins in E.coli using a pBluescrips vector." Gene Expression. 7. 149-161 (998)
[Publications] Rosenbusch,J.P.: "Transmembrane signalling scross ligand-gated FhuA-receptor:crystal structures of free and ferrichrome-bund states reveal allosteric changes." Cell. 95. 771-778 (1998)
[Publications] Rosenbusch,J.P.: "Detergent binding in trigonal crystals of OmpF porin from E.coli." Biochimie. 80. 543-551 (1998)
[Publications] Rosenbusch,J.P: "Stability of trimeric porin:the contributions of the latching loop L2." Biochemistry. 37. 15663-15670 (1998)
[Publications] Rosenbusch,J.P.: "Inactivation in vitro of the Escherichia coli outer membrane protein FhuA by a phage T5-encoded lipoprotein." FEMS Microbiol.Lett.168. 119-125 (1998)
[Publications] Rosenbusch,J.P.: "Assessing the functionality of a membrane protein a three-dimensional crystal." J.Mol.Biol.281. 587-592 (1998)
[Publications] Schirmer,T: "General and specific porins from bacterial outer membranes." J.Struct.Biol. 121. 101-109 (1998)
[Publications] 中江太治編集: "生物学と医学をつなぐ分子細胞生物学" 300 (1998)

1998 Fiscal Year Annual Research Report

抗生物質に特異性を有する透過チャンネルの原子構造と機能

Principal Investigator

中江 太治 東海大学, 医学部, 教授 (50102851)

Research Products

[Publications] Nakae,T.: "Subunit Swapping in the Mex-Extrusion Pumps in Pseudomonas aeruginosa" BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS. 244. 898-902 (1998)

[Publications] 中江太治: "緑膿菌の多剤交叉耐性と薬剤排出" 現代医療. 30. 1249-1254 (1998)

[Publications] Taiji NAKAE,: "Flocculation and Membrane Binding of Outer Membrane Protein F,Porin,at Acidic pH" Jpn.J.Appl.Phys. 37. 2074-2077 (1998)

[Publications] Taiji Nakae: "Identification of the Catalytic Triad of the protein D2 Protease in Pseudomonas aeruginosa" BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS. 247. 142-145 (1998)

[Publications] Taiji Nakae: "Antibiotic stress induces a large amount of outer membrane protein in Pseudomonas aeruginosa" FEMS Microbiology Letters. 165. 261-265 (1998)

[Publications] 中江太治: "非特異的抗生物質耐性の機序" Clinical View. 4. 4-5 (1998)

[Publications] Rosenbusch,J.P: "Coupling site-directed mutagenesis with high-level expression:large scale production of mutant porins from Escherichia coli." FEMS Microbiology Letters. 163. 65-72 (1998)

[Publications] Rosenbusch,J.P.: "Lipidic Cubic Phases:New Matrices for the Three-Dimensional Crystallization of Membrane Proteins." J.Struct.Biol.121. 82-91 (1998)

[Publications] Schirmer,T.: "Crystal structure of the IIB subunit of a fructose permease(IIBLev)from Bacillus subtilis." J.Mol.Biol.276. 591-602 (1998)

[Publications] Schirmer,T: "Crystal structure of human recombinant omithine aminotransferase." J.Mol.Biol. 277. 81-102 (1998)

[Publications] Rummel,G.,Schirmer,T: "Detergent binding in trigonal crystals of OmpF porin from Escherichia coli." Biochimie. 80. 543-551 (1998)

[Publications] Schirmer,T: "Stability of trimeric OmpF porin:the contributions of the latching loop L2" Biochemistry. 37. 15663-15670 (1998)

[Publications] Rosenbusch,J.P.: "Overexpression of outer membrane proteins in E.coli using a pBluescrips vector." Gene Expression. 7. 149-161 (998)

[Publications] Rosenbusch,J.P.: "Transmembrane signalling scross ligand-gated FhuA-receptor:crystal structures of free and ferrichrome-bund states reveal allosteric changes." Cell. 95. 771-778 (1998)

[Publications] Rosenbusch,J.P.: "Detergent binding in trigonal crystals of OmpF porin from E.coli." Biochimie. 80. 543-551 (1998)

[Publications] Rosenbusch,J.P: "Stability of trimeric porin:the contributions of the latching loop L2." Biochemistry. 37. 15663-15670 (1998)

[Publications] Rosenbusch,J.P.: "Inactivation in vitro of the Escherichia coli outer membrane protein FhuA by a phage T5-encoded lipoprotein." FEMS Microbiol.Lett.168. 119-125 (1998)

[Publications] Rosenbusch,J.P.: "Assessing the functionality of a membrane protein a three-dimensional crystal." J.Mol.Biol.281. 587-592 (1998)

[Publications] Schirmer,T: "General and specific porins from bacterial outer membranes." J.Struct.Biol. 121. 101-109 (1998)

[Publications] 中江太治 編集: "生物学と医学をつなぐ分子細胞生物学" 300 (1998)

中江太治東海大学, 医学部, 教授 (50102851)

[Publications] 中江太治編集: "生物学と医学をつなぐ分子細胞生物学" 300 (1998)