Reaction of phenylglyoxal with chicken gizzard myosin.
Modification of chicken gizzard myosin with phenyl[2-14C]-glyoxal inhibited the K+-ATPase (ATP phosphohydrolase, EC 3.6.1.32) activity as a function of time. During the 2.5 and 15 min interval 3.2 mol of the reagent were incorporated per 4.7 X 10(5) g protein and the K+-ATPase activity was 50% inhibited. Phenylglyoxal reacted with arginine residues of gizzard myosin in a mol ratio of two to one, phenylglyoxal to arginine as determined spectrophotometrically. The modification was limited to the subfragment 1 heavy chain and rod-like regions and none of the light chains were lost. The inhibition of the ATPase activity occurred when the subfragment 1 region was modified predominantly. The same results were obtained when the myosin was phosphorylated and then incubated with phenylglyoxal. Substrate MgATP2- or MgADP enhanced the inactivation of gizzard myosin; there was an increase in the incorporation of the reagent and a change in the distribution into the heavy chains. Approx. 0.5 mol of the nucleotide was bound to 4.7 X 10(5) g of phenylglyoxal myosin. Conformational changes, induced by these modifications, were responsible for the inhibition of enzymic activity. Arginine residues of gizzard myosin are necessary for the maintenance of the ATPase activity of this contractile protein.
Bailin
G
G
D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000447:Aldehydes / Q000494:pharmacology*; D000818:Animals; D001120:Arginine; D001665:Binding Sites; D027682:Cation Transport Proteins; D002645:Chickens; D005895:Gizzard, Avian / Q000378:metabolism; D066298:In Vitro Techniques; D009218:Myosins / Q000378:metabolism*; D010658:Phenylglyoxal / Q000378:metabolism / Q000494:pharmacology*; D011487:Protein Conformation
D000447:Aldehydes; D027682:Cation Transport Proteins; D000255:Adenosine Triphosphate; D001120:Arginine; D000251:Adenosine Triphosphatases; C022761:potassium transporting ATPase; D009218:Myosins; D010658:Phenylglyoxal