FEBS Letters | |
Troponin replacement in permeabilized cardiac muscle Reversible extraction of troponin I by incubation with vanadate | |
Strauss, John D.1  Troschka, Monika1  Bletz, Christel1  Zeugner, Claudia1  Rüegg, J.Caspar1  Van Eyk, Jennifer E.1  | |
[1] II Physiologisches Institut, Universität Heidelberg, INF 326, 6900 Heidelberg 1, Germany | |
关键词: Troponin; Troponin I; Vanadate; Contractility; Muscle regulation; TnI; troponin; TnC; troponin C; Tnt; troponin T; DOC-TCA; deoxycholate trichloroacetic acid; ATP; adenosine triphosphate; EDTA; ethylene-diamine tetraacetic acid; EGTA; ethylene glycol (bis(β-aminoethyl ether) N; N; N″; N″ tetraacetic acid; DTE; dithioerythritol; NADH; nicotinamide adenine dinucleotide; UV; ultraviolet; HRP; horseradish peroxidase; | |
DOI : 10.1016/0014-5793(92)81338-M | |
学科分类:生物化学/生物物理 | |
来源: John Wiley & Sons Ltd. | |
【 摘 要 】
Calcium-dependent regulation of tension and ATPase activity in permeabilized porcine ventricular muscle was lost after incubation with 10 mM vanadate. After transfer from vanadate to a vanadate-free, low-Ca2+ solution (pCa > 8), the permeabilized muscle produced 84.8% ± 20.1% (± S.D., n=98) of the isometric force elicited by high Ca22+ (pCa ∼4.5 prior to incubation with vanadate. Transfer back to a high Ca2+ solution elicited no additional force (83.2% ± 18.7% of control force). SDS-PAGE and immunoblot analysis of fibers and solutions demonstrated substantial extraction (>90%) of Troponin I (TnI). Calcium dependence was restored after incubation with solutions containing either whole cardiac troponin or a combination of TnI and troponin C subunits. This reversible extraction of troponin directly demonstrates the role of TnI in the regulation of striated muscle contractility and permits specific substitution of the native TnI with exogenously supplied protein.
【 授权许可】
Unknown
【 预 览 】
Files | Size | Format | View |
---|---|---|---|
RO201912020296930ZK.pdf | 689KB | download |