Effects of protein-modifying reagents on an isoenzyme of potato apyrase.
Treatment of an isoenzyme of potato apyrase of high adenosine triphosphatase/adenosine diphosphatase (ATPase/ADPase) ratio with iodine, N-acetylimidazole or tetranitromethane inactivates the ATPase activity of this enzyme faster than its ADPase activity. There was protection by substrates with the two last-named substances. This and the appearance of nitrotyrosine suggests the participation of tyrosyl residues in both enzymic activities of potato apyrase. The participation of thiol groups is excluded by the insensitivity of apyrase to p-chloromercuribenzoate. Also, 2-hydroxy-5-nitrobenzyl bromide or carboxymethylation produce the same rate of inactivation of ATPase and ADPase activities. Substrates protect both activities from inactivation. Hydrogen peroxide and photo-oxidation inactivate ATPase activity faster than ADPase activity. There is no protection by substrates. Analysis of pH effects on V(max.) and K(m) suggest different pK values for the amino acid residues at the ATP and ADP sites.
D000215:Acylation; D000244:Adenosine Diphosphate; D000251:Adenosine Triphosphatases; D000255:Adenosine Triphosphate; D000596:Amino Acids / Q000032:analysis; D001665:Binding Sites; D002729:Chloromercuribenzoates; D002845:Chromatography; D006863:Hydrogen-Ion Concentration; D007093:Imidazoles; D007455:Iodine; D007527:Isoenzymes; D008745:Methylation; D010084:Oxidation-Reduction; D010744:Phosphoric Monoester Hydrolases; D010944:Plants / Q000201:enzymology*; D014443:Tyrosine; D014675:Vegetables
D000596:Amino Acids; D002729:Chloromercuribenzoates; D007093:Imidazoles; D007527:Isoenzymes; D014443:Tyrosine; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D007455:Iodine; D010744:Phosphoric Monoester Hydrolases; D000251:Adenosine Triphosphatases