| Project/Area Number |
61470157
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
物質生物化学
|
|---|
| Research Institution | Institute of Applied Microbiology, University of Tokyo |
|---|
Principal Investigator |
MATSUHASHI Michio (Professor) Institute of Applied Microbiology, University of Tokyo, 応用微生物研究所, 教授 (40013297)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
富田 啓子 東京大学, 応用微生物研究所, 教務職員
TOMITA Keiko (Research Associate) Institute of Applied Microbiology, University of Tokyo
|
|---|
| Project Period (FY) |
1986 – 1987
|
| Project Status |
Completed (Fiscal Year 1987)
|
| Budget Amount *help |
¥5,500,000 (Direct Cost: ¥5,500,000)
Fiscal Year 1987: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1986: ¥4,500,000 (Direct Cost: ¥4,500,000)
|
|---|
| Keywords | Penicllin-binding protein / Crystallography / Methicillin-resistance / MRSA / Induction / Penicillinase / Cell division / ペニシリン誘導 / 大腸菌 / ブドウ球菌 / MRSA |
|---|
| Research Abstract |
The project centered upon the elucidation of promary and three-dimen-sional structures of penicillin-binding proteins (PBPs) which play important roles in the duplication of bacterial cells, and regulation and expression of their functions.
(1) Crystallization and crystallographic studies of penicillin-binding proteins of Escherichia coli.
Higher molecular weight PBP-1B (component gamma) and lower molecular weight PBP-2 (mutant type) were purified in a large amount from the membranes of E. coli and were crystallized. A preliminary X-ray crystallographic work has been performed. The complete determination of three-dimensional structure of the proteins was aimed in order to determine their molecular shape in the membrane and to enable the designing of an effective <beta>-lactam antibiotic.
(2) Cloning and DNA-sequencing of the <beta>-lactam-inducible, <bate>-lactam-resistant PBP gene in methicillin-resistant Staphylococcus aureus (MRSA).
Cloning and base sequencing of the PBP gene of an MRSA strain, which is supposed to be responsible for the -lactam resistance of MRSA indicated that the MRSA-PBP gene has a regulatory region resembling that of a penicillinase and the major portion of the coding frame resembling a PBP gene of E. coli, suggesting the evolution of the protein by fusion of a penicillinase gene and a PBP gene.
(3) Analysis of genes involved in regulation and expression of functions of PBP.
In E. coli, three regions of the chromosome that contain clusters of genes related with the cell growth and division are involved in regulation and expression of the functions of PBP. These regions, mra (2 min on the E. coli chromosome mpa), mrd (15 min) and mre (71min) regions, were extensively analyzed genetically and physically and some portions of them were sequenced.
In Pseudomonas aeruginosa, lower molecular weight PBP-5 was determined to be involved in the induction of a chromosomal penicillinase.
|
