Assembly and kinetic properties of myosin light chain isozymes from fast skeletal muscle.
Myosin from chicken pectoralis muscle consists of isozymes that differ in their alkali light chains. It is possible to isolate alkali 1 (A1) and alkali 2 (A2) homodimers of native myosin by immunoadsorption methods, and to compare their steady-state kinetics as well as their assembly into synthetic filaments under a variety of ionic conditions. Bipolar filaments of the isozymes formed at low salt concentrations had a narrow length distribution and did not differ from controls made from unfractionated myosin. Chicken myosin also assembles into highly homogeneous minifilaments similar to those formed by rabbit myosin in a citrate/Tris buffer. Analytical ultracentrifugation and electron microscopy showed that A1-homodimer, A2-homodimer and unfractionated myosin assembled into 0.3 micron short, bipolar minifilaments, which were indistinguishable from one another in size and shape. The steady-state myosin ATPase activity of the two homodimeric isozymes was identical in K+(EDTA) and Ca2+ assay media. The actomyosin Mg2+ ATPase measured at 25 and 55 mM-KCl (pH 8.0) showed only minor differences in both Vmax and Kapp. Actomyosin activity was also determined for the more homogeneous minifilament preparations of the isozymes and these, as well, produced essentially indistinguishable kinetic parameters. Thus we find no evidence to support the hypothesis that a particular alkali light chain of myosin can affect either the structure of the filaments or the steady-state rate of ATP hydrolysis.
D000205:Actomyosin / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D002645:Chickens; D003599:Cytoskeleton / Q000648:ultrastructure; D007527:Isoenzymes / Q000378:metabolism; D007700:Kinetics; D008854:Microscopy, Electron; D009132:Muscles / Q000032:analysis* / Q000201:enzymology / Q000648:ultrastructure; D009218:Myosins / Q000378:metabolism
D007527:Isoenzymes; D000205:Actomyosin; D000251:Adenosine Triphosphatases; D009218:Myosins