1996 Fiscal Year Final Research Report Summary
Molecular mechanism of Energy conversion on the biomembrane
| Project/Area Number |
06045012
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | University-to-University Cooperative Research |
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| Research Institution | TOKYO Inst.Technology |
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Principal Investigator |
YOSHIDA Masasuke Tokyo Institule of Technology, Professor, 資源化学研究所, 教授 (90049073)
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| Co-Investigator(Kenkyū-buntansha) |
MONTAL Mauricio Univ.of California, SanDiego, Prof., Univ. of California・San D, 教授
MUNEYUKI Eiro Tokyo Inst.Technology.Asst.Prof., 資源化学研究所, 助手 (80219865)
ALLISON William S Univ.of california, SanDiego, Prof., Univ., 教授
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| Project Period (FY) |
1994 – 1996
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| Keywords | ATPase / F1-ATPase / FoF1-ATPase / ATP synthase / H^+-ATP synthase / Reconstitutioin / Single-site catalysis / Chase-promotioin |
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| Research Abstract |
In order to know how many functional catalytic sites are necessary for ATPase activity of F1-ATPase from a thermophilic Bacillus PS3, a new method to isolate homogeneous prepartion of the alpha3beta3gamma complex with 1,2, or 3 incompetent catalytic sites was developed. Ten glutamic acids (Glu・Tag) were linked to C-terminus of the catalytically incompetent beta (E190Q) subunit. Glu・Tag itself did not affect ATPase activity of the complexes. Two kinds of alpha3beta3gamma complexes, one containing beta (wild-type) and the other Glu・Tag-linked beta (E190Q), were mixed, urea-denatured, dialyzed, and alpha3beta3gamma complexes were reconstituted. Each of the complexes containing different number of Glu・Tag-linked beta (E190Q) was separated by anion-exchange chromatography and analyzed. The results were as follows. 1) Normal steady-state ATPase activity requires three intact catalytic sites. 2) Chase-acceleration, a catalytic cooperativity, requires at least two intact catalytic sites. 3) Single-site catalysis can be mediated by a single intact catalytic site alone. Re-scrambling of subunits between complexes could occur when the complex was aged under some condition and this might be one of the reasons of the previous contradictory result (Miwa et al. (1989) J.Biochem. (Tokyo) 106,730-734).
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[Publications] Amano,T.,Hisabori,T.,Muneyuki,E.,Yoshida,M.: "Catalytic activity of α_3β_3γ complexes of F_1-ATPase with 1,2,or 3 catalytic sites" J.Biol.Chem.271. 17-17, 343-348 (′96)
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「研究成果報告書概要(和文)」より
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[Publications] Matsui, T., Muneyuki, E., Honda, M., Dou, C., Allison, W.S., Yoshida, M: "Catalytic activity of the alpha_3beta_3gamma complex of F_1-ATPase without noncatalytic ncleotide bindirg site" J.Biol.Chem.271(in press). (1997)
Description
「研究成果報告書概要(欧文)」より
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