Solubilization and partial purification of protein kinase systems from brain membranes that phosphorylate calspectin

FEBS Letters

Sobue, Kenji¹  Kanda, Keiko¹  Kakiuchi, Shiro¹ 

[1] Department of Neurochemistry and Neuropharmacology, Institute of Higher Nervous Activity, Osaka University Medical School, Nakanoshima, Kita-ku, Osaka 530, Japan

关键词: Calspectin;    Fodrin;    Calmodulin-binding protein;    Calmodulin-dependent protein kinase;    EGTA;    ethyleneglycol bis(β-aminoethylether)-N;    N;    N'-tetraacetic acid;    EDTA;    ethylenediamine tetraacetic acid;    SDS;    sodium dodecyl sulfate;    PMSF;    phenylmethylsulfonyl fluoride;    DFP;    diisopropylfluorophosphate;    M r;    relative molecular mass;   

DOI  :  10.1016/0014-5793(82)81331-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

In brain tissue a spectrin-like calmodulin-binding protein calspectin, or fodrin, is concentrated in a synaptosome fraction, where most of the calspectin is associated with the synaptic membranes. This endogenous calspectin was phosphorylated by protein kinase system(s) associated with the membranes. Here, we report the solubilization and partial purification of the membrane-associated calspectin kinase activity. The activity was resolved on a gel filtration column into two fractions, peaks I and II having estimated M _r of 800 000 and 88 000. The activity of peak I was dependent on the presence of both Ca²⁺ and calmodulin. Peak II revealed a basal activity in the absence of Ca²⁺ and calmodulin, which was stimulated 2-fold by addition of Ca²⁺. Calmodulin had no effect on the peak II activity.

【 授权许可】

Unknown