MUTAGENESIS OF THR-286 IN MONOMERIC CA2+/CALMODULIN-DEPENDENT PROTEIN KINASE-II ELIMINATES CA2+/CALMODULIN-INDEPENDENT ACTIVITY

被引：67

作者：

WAXHAM, MN ^{[1
]}

ARONOWSKI, J ^{[1
]}

WESTGATE, SA ^{[1
]}

KELLY, PT ^{[1
]}

机构：

[1] UNIV TEXAS,SCH MED,DEPT NEUROBIOL & ANAT,HOUSTON,TX 77225

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 1990年 / 87卷 / 04期

关键词：

autophosphorylation; bacterial expression; pseudosubstrate;

D O I：

10.1073/pnas.87.4.1273

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

We have examined the role of Thr-286 autophosphorylation in the autoregulation of Ca2+/calmodulin-dependent protein kinase II. Using site-directed mutagenesis, we have substituted alanine or serine for Thr-286, or isoleucine for Arg-283, in the 50-kDa subunit of the kinase and expressed each protein in bacteria. Activation and autophosphorylation of all four enzymes were stringently dependent on Ca2+/calmodulin, indicating that neither Arg-283 nor Thr-286 is an absolute requirement for the pseudosubstrate inhibition of the enzyme. Autophosphorylation of the Ile-283 or Ala-286 enzyme generated little, if any, Ca2+/calmodulin-independent kinase activity, unlike the parent (Thr-286) or Ser-286 enzyme. The enzymes expressed in bacteria are predominantly monomeric, indicating that the generation of Ca2+/calmodulin-independent activity does not require the cooperative interactions of subunits normally present in the brain holoenzyme.

引用

页码：1273 / 1277

页数：5

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