| Project/Area Number |
25670276
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Laboratory medicine
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| Research Institution | Nara Medical University |
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Principal Investigator |
YAMAMOTO Keizo 奈良県立医科大学, 医学部, 准教授 (90254490)
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| Co-Investigator(Kenkyū-buntansha) |
KASAHARA Kei 奈良県立医科大学, 医学部, 准教授 (50405403)
NAKAYAMA Akifumi 岐阜医療科学大学, 保険科学部, 教授 (70536721)
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| Project Period (FY) |
2013-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2014: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Keywords | βーラクタマーゼ / X線結晶構造解析 / 基質特異性 / 構造変化 / β―ラクタマーゼ / Ｘ線結晶構造解析 / 多剤耐性 / タンパク質精製 / クローニング / 大量発現 / 大腸菌 |
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| Outline of Final Research Achievements |
Plasmid-mediated extended spectrum beta-lactamases (ESBLs) named CTX-M causes hospital- and community-acquired infections in Japan. Comparison of DNA sequences and structures of CTX-Ms showed that amino acid substitution in the three loop regions, omega-loop, VNYNP-loop, and a loop connecting alpha/beta domain mast be important to change of substrate specificity. Ala-219 which locates in a loop connecting alpha/beta domain is considered as a key residue to substrate specificity. Thus, the structures of three mutant enzymes, A219V, A219L, A219F, and wild type CTX-M-2 were solved. Although the structural difference among wild type, A219V, and A219F were very small, maximum XYZ-difference between wild type and A219L were 0.42 nm. It reveals that the structure of CTX-Ms is latently flexible and mutation in the loop region raises changes of overall structure and substrate specificity.
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