ISOLATION OF CARDIAC MYOFIBRILS AND MYOSIN LIGHT-CHAINS WITH INVIVO LEVELS OF LIGHT CHAIN PHOSPHORYLATION

被引：81

作者：

HOLROYDE, MJ

SMALL, DAP

HOWE, E

SOLARO, RJ

机构：

[1] UNIV CINCINNATI,COLL MED,DEPT PHYSIOL,CINCINNATI,OH 45267

[2] UNIV CINCINNATI,DEPT PHARMACOL & CELL BIOPHYS,CINCINNATI,OH 45267

来源：

BIOCHIMICA ET BIOPHYSICA ACTA | 1979年 / 587卷 / 04期

基金：

美国国家卫生研究院;

关键词：

(Heart myofibril); Catecholamine; Light chain; Myosin; Phosphorylation;

D O I：

10.1016/0304-4165(79)90014-X

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Conditions are described for the preparation of functional myofibrils and myosin light chains from freeze-clamped beating hearts with the state of light chain phosphorylation chemically 'frozen' during the extraction procedure. Myofibrils were shown to be functionally intact by measurement of Ca2+ binding and ATPase activity. Highly purified cardiac myosin light chains could be routinely isolated from myofibrillar preparations using ethanol fractionation together with ion-exchange chromotography. Analysis of light chains for covalent phosphate indicated that basal levels of phosphorylation of the 18-20 000 dalton light chain of myosin in rabbit hearts beating in situ or in a perfusion apparatus were 0.3-0.4 mol/mol. Covalent phosphate content of the light chain fraction did not change during perfusion of hearts with 10 μM epinephrine. © 1979.

引用

页码：628 / 637

页数：10

共 24 条

[1]

BAGSHAW C R, 1977, Biochemistry, V16, P59, DOI 10.1021/bi00620a010

[2]

BARANY K, 1977, J BIOL CHEM, V252, P4752

[3] ISOLATION AND PROPERTIES OF PLATELET MYOSIN LIGHT CHAIN KINASE [J].