1994 年度実績報告書

薬剤耐性に係わる細胞膜の機能及びそれと連係的に作用する因子の総合的研究

研究課題

研究課題/領域番号	05304028
研究機関	東海大学
研究代表者	中江太治東海大学, 医学部, 教授 (50102851)
研究分担者	平松啓一順天堂大学, 医学部, 教授 (10173262) 後藤直正京都薬科大学, 薬学部, 講師 (30121560) 山口明人千葉大学, 薬学部, 助教授 (60114336) 井上松久北里大学, 医学部, 教授 (10008336) 伊豫部志津子群馬大学, 医学部, 助教授 (90008318)
キーワード	楽剤耐性 / 細菌感染 / 多剤耐性 / 細胞膜 / 排出 / 病院内感染 / MRSA / 透過性
研究概要	本年度は大きく分けて4本の研究テーマを柱として共同研究を行った。(i)グラム陰性菌の外膜が薬剤耐性に果たす役割。中江は外膜蛋白質OprD2蛋白質がイミペネムに対して特異性を有することを平面脂質二重膜法を用いて単分子レベルで明らかとした。後藤は0prD2に対する単クローン抗体を用いて0prD2の細胞外に突出しているドメインの解析をおこなった。良原は0prD2にプロテアーゼ活性があることを発見した。(ii)内膜における薬剤排出による耐性。中江は緑膿菌における薬剤排出に係わる遺伝子をクローン化し、その塩基配列を調べたところ、それは鉄のキレ-夕であるpyoverdineの輸送蛋白質と同一であることを明らかとした。山口はテトラサイクリン排出蛋白におけるテトラサイクリン排出に必須のアミノ酸残基を決定する実験を行った。その結果必須領域Asp282に変異を挿入するとAla220が酸性アミノ酸に変異することなどからアミノ酸残基285-220位を含む領域に正味の酸性アミノ酸残基が必須であることを明らかとした。笹津はブドウ球菌球菌における消毒剤耐性遺伝子はすべてのブドウ球菌に存在することを明らかとした。(iii)MRSAと細胞膜、平松はMRSAのメチシリン耐性を担うmecA遺伝子の全塩基配列を決定した。このクローンをプローブとして種々の菌のDNAをサザン方で解析したところmec領域DNAが場所特異的組み替えによって挿入されたことの確証を得た。神尾はMRSAのへモリジンが作用するにはF成分の初発の膜への結合が引き金となってつぎにHγllが作用することを明らかとした。(iv)細胞膜と他の因子による連係と耐性の係わり。伊豫部は臨床分離株における新キノロン剤高度耐性はnfxBやnfxC変異とDNAgyrase(nfxA)の二重変異によることを明らかとした。井上はβラクタマーゼの誘導にPBPの代謝産物が係わっている可能性を示した。以上述べたように本総合研究は予定通りに進行している。

研究成果
(26件)

すべてその他

すべて文献書誌 (26件)

[文献書誌] Inoue,M.Nonoyama,R.,Okamoto,R.and Ida,T.: "Antimicrobial activity aminoglycoside antibiotics against methicllin-resistant S.aureus" Drugs Exptl.Cli.Res.20. 233-240 (1994)
[文献書誌] Hayashi,I.,Inoue,M.and Hashimoto,H: "Nationwide investigation in Japan on the efficacy of arbekacin in MASA infection" Drugs Exptl.Cli.Res. 20. 225-232 (1994)
[文献書誌] Ozawa,T.,Kaneko,J.and Kamio,Y.: "Inactivation of γ-hemolusin HγII component by addition of monosialoganglioside GM1 to human erythrocyte" Biosci.Biotech.Biochem.58. 602-605 (1994)
[文献書誌] Choorit,W.and Kamio,Y.: "Existence of a new protein component with the same function as the LukF component of leukocidin or γ-hemolysin and its gene in Staphylococcus aureus P83" FEBS Lett.375. 260-264 (1995)
[文献書誌] Iyobe,S.and Mitsuhashi,S.: "Cloning and expression in Enterobacteriaceae of the extended-spectrum β-lactamase gene from a Pseudomonas aerigompsa plasmid" FEMS Microbiol.Lett.121. 175-180 (1994)
[文献書誌] Yoshida,T.,Iyobe,S.and Mitsuhashi,S.: "Mechanism of high-lebvel resistance to quinolones in urinary tract isolates of Pseudomonas aeruginosa" Antimicrob.Agents.Chemother.38. 1466-1469 (1994)
[文献書誌] Yokota,S.and Matsumoto,H.: "Identification of the Lipopolysaccharide core region as the receptor site for a Cytotoxin-converting phage φCTX of Pseudomonas aeruginosa" J.Bacteriology.176. 5262-5269 (1994)
[文献書誌] Hayashi,T.,Matsumoto,H.and Tcrawaki,Y.: "Cytotoxin-converting phages,φCTX and PS21,are R pyocin-related phages" FEMS Microbiol.Lett.122. 239-244 (1994)
[文献書誌] Kawakami,Y.and Matsumoto,H.: "Restriction fragment length polymorphism (RFLP)of genomic DNA of moraxella(Branhamella)catarrhalis isolates in a Hospital" Microbiol.Immunol.38. 891-895 (1994)
[文献書誌] Kawakami,Y.and Matsumoto,H.: "In vitro activity of arbekacin against clinical isolates of methicillin-resistant Staphylococcus aureus in a hospital" J.Antibiotics.47. 507-509 (1994)
[文献書誌] Ito,T.,Hiramatsu,K.and Yokota,T.: "Identification of a novel sugar,2-0-methyl-4-amino-4.6-dideoxy mannose in the.Iipopolysaccharide of V.cholerae" Carbohydrate Research.256. 113-128 (1994)
[文献書誌] Someya,y.and Yamaguchi,A.: "Site-specificity of the second-site suppressor mutation of the Asp285->Asn mutant of Metal-tetracycline/H^+antiporter pf Escherichia coli and the effects of amino acid substitutions at the first and second sites" Biochemistry.34. 7-12 (1995)
[文献書誌] Someya,Y.,Yamaguchi,A.and Sawai,T.: "A novel glycylcycline,9-(N,N-dimethylglycylamido)-6-demethyl-6-deoxytetracycline,is neither transported nor recognized by the transposon Tn10-encoded metal-tetracycline/H^+" Antimicrob.Agents.Chemother.39. 247-249 (1995)
[文献書誌] Yamaguchi,A.,Kimura,T.and Sawai,T.: "Hot spot for sulfhydry inactivation of Cys mutants in the widely conserver sequence motifs of the meta-tetracycline/H^+antiporter of Escherichia coli" J.Biochem.115. 958-964 (1994)
[文献書誌] Sato,K.,Yamaguchi,A.and Yoshida,M.: "Tetracycline/H^+antiporter was degraded repidly in Escherichia coli cells when truncated at last transmembrane helix and this degradation was protected by overproduced GroEL/FS" Biochem.Biophys.Res.Commun.202. 258-264 (1994)
[文献書誌] Sasatsu.,M.and Kono,M: "Evaluation of antiseptics by the modified phenol coefficient method:sensitivity of methcillin-resistant Staphylococcus aureus to antiseptics" Biol.Pharm.Bull.17. 136-138 (1994)
[文献書誌] Sasatsu.,M.and Kono,M: "Substrates and inhibitors ot antiseptics resistance in Staphylococcus aureus" Biol.Pham.Bull.17. 163-165 (1994)
[文献書誌] Noguchi,N.,Sasatsu,M.,and Kono,M.: "Expression in Escherichia coli of a TetK determinant from Staphylococcus aureus" Biol.Pharm.Bull.17. 352-355 (1994)
[文献書誌] Sasatus,M.,and Kono,M.: "Effect of β-lactamase on minimum inhibitory concentrations of methicillin in methicillin-resistant Staphylococcus aureus" Microbios.78. 145-153 (1994)
[文献書誌] Gotoh,N.and,D.E.: "Isolation and characterization of the outer membrane proteins of Burkholderia(Pseudomonas)pseudomallei" Microbiology.140. 797-805 (1994)
[文献書誌] Gotoh,N.and Nishino,T.: "Isolation of OprM-deficient mutants of Pseudomonas aeruqinosa by transposon insertion mutagenesis:Evidence of involvement in multiple antibitic resistance" FEMS Microbiol.Lett.122. 267-274 (1994)
[文献書誌] Gotoh,N.and Nishino,T.: "Evidence confirming the location of OprM in the Pseudomonas aeruginosa outer membrane" FEMS Microbiol.Lett.122. 309-312 (1994)
[文献書誌] Gotoh,N.and Nishino,T.: "Burkholderia(formerly Pseudomonas)cepaciaporins is an oligomer of two component proteins" Microbiology.140. 3285-3291 (1994)
[文献書誌] Hori,S.,Hiramatsu,K.and Yokota,T.: "Staphylococci and Staphylococcal infections,(Sequential acquisition of newquinolone resistance by methicillin-resistant and susceptible Staphylococcus aureus)" 3 (1994)
[文献書誌] Hiramatsu,K.and Yokota,T.: "Staphylococci and Staphylococcal infections,(Methicillin resistance regulation of the prototype MRSA strain.)" 3 (1994)
[文献書誌] Suzuki,E.and Hiramatsu,K.: "Recent Advances in Chemother,(Comparative study of in vitro antimicrobial activity of a newly synthesized quinolone,AM-1155,and other 4-quiinolone antibiotics)" 2 (1994)

1994 年度 実績報告書

薬剤耐性に係わる細胞膜の機能及びそれと連係的に作用する因子の総合的研究

研究代表者

中江 太治 東海大学, 医学部, 教授 (50102851)

研究成果

[文献書誌] Inoue,M.Nonoyama,R.,Okamoto,R.and Ida,T.: "Antimicrobial activity aminoglycoside antibiotics against methicllin-resistant S.aureus" Drugs Exptl.Cli.Res.20. 233-240 (1994)

[文献書誌] Hayashi,I.,Inoue,M.and Hashimoto,H: "Nationwide investigation in Japan on the efficacy of arbekacin in MASA infection" Drugs Exptl.Cli.Res. 20. 225-232 (1994)

[文献書誌] Ozawa,T.,Kaneko,J.and Kamio,Y.: "Inactivation of γ-hemolusin HγII component by addition of monosialoganglioside GM1 to human erythrocyte" Biosci.Biotech.Biochem.58. 602-605 (1994)

[文献書誌] Choorit,W.and Kamio,Y.: "Existence of a new protein component with the same function as the LukF component of leukocidin or γ-hemolysin and its gene in Staphylococcus aureus P83" FEBS Lett.375. 260-264 (1995)

[文献書誌] Iyobe,S.and Mitsuhashi,S.: "Cloning and expression in Enterobacteriaceae of the extended-spectrum β-lactamase gene from a Pseudomonas aerigompsa plasmid" FEMS Microbiol.Lett.121. 175-180 (1994)

[文献書誌] Yoshida,T.,Iyobe,S.and Mitsuhashi,S.: "Mechanism of high-lebvel resistance to quinolones in urinary tract isolates of Pseudomonas aeruginosa" Antimicrob.Agents.Chemother.38. 1466-1469 (1994)

[文献書誌] Yokota,S.and Matsumoto,H.: "Identification of the Lipopolysaccharide core region as the receptor site for a Cytotoxin-converting phage φCTX of Pseudomonas aeruginosa" J.Bacteriology.176. 5262-5269 (1994)

[文献書誌] Hayashi,T.,Matsumoto,H.and Tcrawaki,Y.: "Cytotoxin-converting phages,φCTX and PS21,are R pyocin-related phages" FEMS Microbiol.Lett.122. 239-244 (1994)

[文献書誌] Kawakami,Y.and Matsumoto,H.: "Restriction fragment length polymorphism (RFLP)of genomic DNA of moraxella(Branhamella)catarrhalis isolates in a Hospital" Microbiol.Immunol.38. 891-895 (1994)

[文献書誌] Kawakami,Y.and Matsumoto,H.: "In vitro activity of arbekacin against clinical isolates of methicillin-resistant Staphylococcus aureus in a hospital" J.Antibiotics.47. 507-509 (1994)

[文献書誌] Ito,T.,Hiramatsu,K.and Yokota,T.: "Identification of a novel sugar,2-0-methyl-4-amino-4.6-dideoxy mannose in the.Iipopolysaccharide of V.cholerae" Carbohydrate Research.256. 113-128 (1994)

[文献書誌] Someya,y.and Yamaguchi,A.: "Site-specificity of the second-site suppressor mutation of the Asp285->Asn mutant of Metal-tetracycline/H^+antiporter pf Escherichia coli and the effects of amino acid substitutions at the first and second sites" Biochemistry.34. 7-12 (1995)

[文献書誌] Someya,Y.,Yamaguchi,A.and Sawai,T.: "A novel glycylcycline,9-(N,N-dimethylglycylamido)-6-demethyl-6-deoxytetracycline,is neither transported nor recognized by the transposon Tn10-encoded metal-tetracycline/H^+" Antimicrob.Agents.Chemother.39. 247-249 (1995)

[文献書誌] Yamaguchi,A.,Kimura,T.and Sawai,T.: "Hot spot for sulfhydry inactivation of Cys mutants in the widely conserver sequence motifs of the meta-tetracycline/H^+antiporter of Escherichia coli" J.Biochem.115. 958-964 (1994)

[文献書誌] Sato,K.,Yamaguchi,A.and Yoshida,M.: "Tetracycline/H^+antiporter was degraded repidly in Escherichia coli cells when truncated at last transmembrane helix and this degradation was protected by overproduced GroEL/FS" Biochem.Biophys.Res.Commun.202. 258-264 (1994)

[文献書誌] Sasatsu.,M.and Kono,M: "Evaluation of antiseptics by the modified phenol coefficient method:sensitivity of methcillin-resistant Staphylococcus aureus to antiseptics" Biol.Pharm.Bull.17. 136-138 (1994)

[文献書誌] Sasatsu.,M.and Kono,M: "Substrates and inhibitors ot antiseptics resistance in Staphylococcus aureus" Biol.Pham.Bull.17. 163-165 (1994)

[文献書誌] Noguchi,N.,Sasatsu,M.,and Kono,M.: "Expression in Escherichia coli of a TetK determinant from Staphylococcus aureus" Biol.Pharm.Bull.17. 352-355 (1994)

[文献書誌] Sasatus,M.,and Kono,M.: "Effect of β-lactamase on minimum inhibitory concentrations of methicillin in methicillin-resistant Staphylococcus aureus" Microbios.78. 145-153 (1994)

[文献書誌] Gotoh,N.and,D.E.: "Isolation and characterization of the outer membrane proteins of Burkholderia(Pseudomonas)pseudomallei" Microbiology.140. 797-805 (1994)

[文献書誌] Gotoh,N.and Nishino,T.: "Isolation of OprM-deficient mutants of Pseudomonas aeruqinosa by transposon insertion mutagenesis:Evidence of involvement in multiple antibitic resistance" FEMS Microbiol.Lett.122. 267-274 (1994)

[文献書誌] Gotoh,N.and Nishino,T.: "Evidence confirming the location of OprM in the Pseudomonas aeruginosa outer membrane" FEMS Microbiol.Lett.122. 309-312 (1994)

[文献書誌] Gotoh,N.and Nishino,T.: "Burkholderia(formerly Pseudomonas)cepaciaporins is an oligomer of two component proteins" Microbiology.140. 3285-3291 (1994)

[文献書誌] Hori,S.,Hiramatsu,K.and Yokota,T.: "Staphylococci and Staphylococcal infections,(Sequential acquisition of newquinolone resistance by methicillin-resistant and susceptible Staphylococcus aureus)" 3 (1994)

[文献書誌] Hiramatsu,K.and Yokota,T.: "Staphylococci and Staphylococcal infections,(Methicillin resistance regulation of the prototype MRSA strain.)" 3 (1994)

[文献書誌] Suzuki,E.and Hiramatsu,K.: "Recent Advances in Chemother,(Comparative study of in vitro antimicrobial activity of a newly synthesized quinolone,AM-1155,and other 4-quiinolone antibiotics)" 2 (1994)

1994 年度実績報告書

中江太治東海大学, 医学部, 教授 (50102851)