Phosphorylation of thymus myosin increases its apparent affinity for actin but not its maximum adenosinetriphosphatase rate.
Vertebrate nonmuscle myosins contain two phosphorylatable light chains. The maximum rate, Vmax, of the actin-activated adenosinetriphosphatase (ATPase) of unphosphorylated calf thymus myosin was found to be about 100 nmol/(min X mg), the same as that of thymus myosin with two phosphorylated light chains. However, the Kapp (actin concentration required to achieve 1/2 Vmax) of the unphosphorylated myosin was 15-20-fold greater than that of the phosphorylated myosin. When actin complexed with either skeletal muscle tropomyosin or calf thymus tropomyosin was used, the values for Vmax were about the same as those obtained with F-actin. In the presence of skeletal muscle tropomyosin, the Kapp of the unphosphorylated myosin was only 2-3-fold greater than that of the phosphorylated myosin, and in the presence of thymus tropomyosin, there was about a 5-fold difference in their Kapp values. Thus, light chain phosphorylation regulates the actin-activated ATPase of thymus myosin not by increasing Vmax but rather by decreasing the Kapp of this myosin for actin. These rather small differences in Kapp suggest that other proteins may be involved in the regulation of the actin-activated ATPase of thymus myosin. Regulated actin (actin plus skeletal muscle troponin-tropomyosin) was used to examine possible effects of thin-filament regulatory proteins. In the presence of calcium, phosphorylation caused only a slight increase in Vmax and a 2-fold decrease in Kapp of the regulated actin-activated ATPase of thymus myosin.(ABSTRACT TRUNCATED AT 250 WORDS)
D000199:Actins / Q000378:metabolism*; D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D004789:Enzyme Activation; D005895:Gizzard, Avian / Q000378:metabolism; D007700:Kinetics; D008970:Molecular Weight; D009132:Muscles / Q000378:metabolism; D009218:Myosins / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D011817:Rabbits; D013950:Thymus Gland / Q000201:enzymology*; D014335:Tropomyosin / Q000378:metabolism; D014422:Turkeys
D000199:Actins; D014335:Tropomyosin; D000251:Adenosine Triphosphatases; D009218:Myosins