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標題: Calcium-dependent protein-protein interactions induce changes in proximity relationships of Cys48 and Cys64 in chicken skeletal troponin I
作者: Liou, Y.M.
Chen, M.W.
關鍵字: proximity relationships;cysteine residues;pyrene-containing;fluorophores;monomer fluorescence;excimer fluorescence;amino-acid sequence;pyrene excimer fluorescence;biological-activity;regulatory domain;proteolytic fragments;angstrom resolution;muscle-contraction;inhibitory region;thin-filaments;cross-linking
Project: European Journal of Biochemistry
期刊/報告no:: European Journal of Biochemistry, Volume 270, Issue 14, Page(s) 3092-3100.
The goal of this study was to relate conformational changes in the N-terminal domain of chicken troponin I (TnI) to Ca2+ activation of the actin-myosin interaction. The two cysteine residues in this region (Cys48 and Cys64) were labeled with two sulfhydryl-reactive pyrene-containing fluorophores [N-(1-pyrene)maleimide, and N-(1-pyrene)iodoacetamide]. The labeled TnI showed a typical fluorescence spectrum: two sharp peaks of monomer fluorescence and a broad peak of excimer fluorescence arising from the formation of an excited dimer (excimer). Results obtained show that forming a binary complex of labeled TnI with skeletal TnC (sTnC) in the absence of Ca2+ decreases the excimer fluorescence, indicating a separation of the two residues. This reduction in excimer fluorescence does not occur when labeled TnI is complexed with cardiac TnC (cTnC). The latter causes only partial activation of the Ca2+-dependent myofibrillar ATPase. The binding of Ca2+ to the two N-terminal sites of sTnC causes a significant decrease in excimer fluorescence and an increase in monomer fluorescence in complexes of labeled TnI with skeletal TnC or TnC/TnT, while Ca2+ binding to site II of cTnC only causes an increase in monomer fluorescence but no change in excimer fluorescence. Thus a conformational change in the N-terminal region of TnI may be necessary for full activation of muscle contraction.
ISSN: 0014-2956
DOI: 10.1046/j.1432-1033.2003.03700.x
Appears in Collections:生命科學系所

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