Characterization of nucleoside diphosphatase in the onion root tip. III. Solubilization and activation of membrane-bound enzyme(s).
The latency of nucleoside diphosphatase (NDPase) in onions root homogenates has been examined by comparing the activation of NDPase activity resulting from detergent treatment with that due to storage of homogenates for several days in the cold. Both detergent treatment and cold storage activated NDPase approximately two-fold. In both cases this activation was paralleled by the loss of enzyme activity from the membrane fractions and its appearance in the supernatants. Electrophoresis of these supernatants revealed an identifical isoenzyme pattern of 5 NDPase bands for both preparations. Enzyme kinetic studies demonstrated that NDPase from the detergent-treated homogenate and the homogenate stored in the cold as well as NDPase from the membrane and supernatant fractions from each of the homogenates all had the same Km value. These data suggest that latency of NDPase is the result of a breakdown of cellular membranes and subsequent release of NDPase.


BACKGROUND
DOC, deoxycholate; NDPase, nucleoside diphosphatase; IDP, inosine 5'-diphosphate; UDP, uridine 5'-diphosphate; GDP, guanosine 5'-diphosphate.
D017766:Acid Anhydride Hydrolases; D003080:Cold Temperature; D003840:Deoxycholic Acid / Q000494:pharmacology; D003902:Detergents / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation / Q000187:drug effects; D007290:Inosine Diphosphate / Q000378:metabolism; D007700:Kinetics; D008566:Membranes / Q000201:enzymology; D010744:Phosphoric Monoester Hydrolases / Q000032:analysis / Q000378:metabolism; D010944:Plants / Q000201:enzymology*; D012995:Solubility
D003902:Detergents; D003840:Deoxycholic Acid; D007290:Inosine Diphosphate; D010744:Phosphoric Monoester Hydrolases; D017766:Acid Anhydride Hydrolases; C021446:inosine diphosphatase