1999 Fiscal Year Final Research Report Summary
Study of the cytoskeleton and lipid biosynthetic system of an archaeon Thermoplasma
| Project/Area Number |
09839033
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
自然史科学
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| Research Institution | TOKYO UNIVERSITY OF PHARMACY AND LIFE SCIENCE |
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Principal Investigator |
YAMAGISHI Akihiko Tokyo University of Pharmasy and Life Science, School of Life Science, Associate professor, 生命科学部, 助教授 (50158086)
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| Co-Investigator(Kenkyū-buntansha) |
ARAKAWA Hideo Tokyo Institute of Technology, Department of Life Science, 生命理工学部, 助手 (80211704)
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| Project Period (FY) |
1997 – 1999
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| Keywords | Thermoplasma / cytoskeleton / Archaea / archaebacteria / etherlipid / terbinafine / tetraetherlipid |
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| Research Abstract |
(1) Cytoskeleton-like structures of Thermoplasma were biochemically analyzed. Cells of Thermoplasma were treated by non-ionic detergent Triton X-100. The solubilized proteins were fractionated by the combination of anion-exchange-column chromatography and gel-exclusion chromatography. Several fractions of proteins that can reversibly polymerize were recovered. Tubular structures and net-like structures were found in some of those fractions by electron microscopic inspection. Peptide composition of the fractions was analyzed by SDS-polyacrylamide gel electophoresis.
(2) Carbon 14-labeled mevalonic acid was used to trace the biosynthetic pathway of ether-membrane lipids in Thermoplasma. Polar ether-lipid fraction of Thermoplasma was extracted from the cells grown in the presence of C14-mevalonic acid. The extracted ether-lipids were analyzed by two-dimensional thin layer chromatography. Lipids were recovered from respective spot of TLC plates and the structures of the lipids were analyzed and estimated. After 2-h incubation, major polar tetraetherlipid, which has phosphatidylglycerol and a sugar molecule, was accumulated in the cells. In the presence of the inhibitor terbinafine, biosynthesis of the major polar lipid was inhibited and a diether lipid was accumulated. After the removal of the inhibitor, biosynthesis of polar lipids was analyzed by thin layer chromatography. The model of the biosynthetic pathway of the tetra-lipids was proposed.
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