| Project/Area Number |
05640646
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
機能・物性・材料
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| Research Institution | Gunma University |
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Principal Investigator |
KOBASHI Harumichi Gunma University ASSOCIATE PROFESSOR, 工学部, 助教授 (80008515)
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| Co-Investigator(Kenkyū-buntansha) |
EIKI Toshio Gunma University ASSOCIATE PROFESSOR, 工学部, 助教授 (60143202)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1994: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1993: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | Myosin / Fluorescent Probe / Noncovalently Bonding Probe / Covalently bonding Prove / SH-Modification / NH_2-Modification / Pyrene Derivatives / Trinitrophenylation / ATPase活性 / ピレンヨードアセトアミド / ピレンメチルヨードアセテイト / ピレンマレイミド / SH基特異的修飾 / NH_2基特異的修飾 / 筋収縮蛋白質 / ピレン / ピレンスルホン酸 / 結合定数 / レーザー光分解 / 蛍光寿命解析 |
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| Research Abstract |
The present research project aimed to elucidate fluorescence behavior of pyrene (P) derivatives which are expected as useful probes for obtaining informations concerning microstructure and functions of M,especially in relation to discrimination of two myosin heads.
1. Binding constants of P and 1-pyrenesulfonate ion (PS) with M were estimated to be 4.9x10^5 and 1x10^7 M^<-1>, respectively, in aq soln (pH 7.0) at 20ﾟC
2. Pyrene molecules of <greater than or equal> 65% fraction were located in inner hydrophobic regions of M where iodide ion as quencher can not quench the fluorescence of P.PS binds with M in a relatively hydrophillic region accessible to the
3. .Some fluorescent probes with a pyrene chromophore react specifically with active SH-or NH_2-groups of M.Myosin modified with N- (1-pyrene) iodoacetamide showed a typical curve in pH dependence of Ca-ATPase activity of SH1-modified M.Change in fluorescence characteristics of the probe was observed associated with conformational change of M induced by the addition of ATP.
4. Trinitrophenylation and retrinitrophenylation with 2,4,6-trinitrobenzenesulfonic acid revealed that the two heads of chicken skeletal M were different from each other in pH dependence of Ca-ATPase activity and in reactivities in the presence and absence of PPi. These findings suport the hypothesis of non-identical heads.
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