Isolation of the calmodulin‐dependent protein kinase system from rabbit skeletal muscle sarcoplasmic reticulum

FEBS Letters

Tuana, Balwant S.¹  MacLennan, David H.² 

[1] Department of Pharmacology, Medical Sciences Building, University of Toronto, Toronto, Ontario M5S 1A8, Canada;The Banting and Best Department of Medical Research, Charles H. Best Institute, University of Toronto, Toronto, Ontario M5G 1L6, Canada

关键词: Protein phosphorylation;    Sarcoplasmic reticulum;    Calmodulin;   

DOI  :  10.1016/0014-5793(88)81266-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A calmodulin-dependent protein kinase system from the sarcoplasmic reticulum was dissolved in Nonidet P40, adsorbed to a CaM affinity column in the presence of Ca²⁺ and eluted in the presence of EGTA. The purified fraction contained major proteins of 60 and 20 kDa and minor components of 89 and 34 kDa, all of which were phosphorylated with dependencies on Ca²⁺, CaM, ATP and pH similar to those observed in the sarcoplasmic reticulum. Differences in the phosphopeptides produced by partial proteolysis of the individual phosphoproteins indicated that they are distinct entities. ¹²⁵I-CaM labeled only the 60 kDa protein, suggesting that it is a kinase.

【 授权许可】

Unknown