Multiple forms of rat liver cysteinesulfinate decarboxylase.
Cysteinesulfinate decarboxylase, purified from male rat livers and homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, is resolved into five distinct enzyme species (isoforms) by gel isoelectric focusing. Since the isoforms are present in fresh liver homogenates and do not arise by proteolysis, the enzyme is apparently heterogeneous in vivo. Although female rat livers contain only 5% of the cysteinesulfinate decarboxylase activity of male livers, immunological and enzymatic studies indicate that the distribution of isoforms is similar in both sexes. Rat brain and kidney also contain multiple isoforms which are cross-reactive with polyclonal antibodies prepared to the liver enzyme. The enzyme exhibits a protomer Mr of 53,000, and the native enzyme is shown by cross-linking studies to be dimeric. Purified enzyme contains no carbohydrate or phosphate and does not bind excess pyridoxal 5'-phosphate. Two pools of enzyme activity are resolved preparatively by chromatofocusing chromatography and have been examined with respect to substrate and inhibitor specificity. Both pools are most active toward L-cysteinesulfinate and L-cysteinesulfonate. Aspartate, homocysteinesulfinate, homocysteinesulfonate, 2-amino-3-phosphonopropionate, and glutamate are decarboxylated at rates less than 1% of that observed with L-cysteinesulfinate; D-cysteinesulfinate is not decarboxylated but is an effective inhibitor. The enzyme isoforms cannot be distinguished on the basis of substrate affinity or specificity. The enzyme is irreversibly inactivated by the mechanism-based inhibitors beta-methylene-DL-aspartate and beta-ethylidene-DL-aspartate. beta-Ethylideneaspartate, in contrast to the beta-methylene derivative, does not inhibit aspartate aminotransferase, an enzyme also important in cysteinesulfinate metabolism. beta-Ethylidene aspartate or related beta-ethylidene compounds may be useful in selectively altering cysteinesulfinate metabolism in vivo.
D000818:Animals; D001921:Brain / Q000201:enzymology; D002262:Carboxy-Lyases / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D005260:Female; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000302:isolation & purification*; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology*; D046911:Macromolecular Substances; D008297:Male; D008970:Molecular Weight; D051381:Rats; D013379:Substrate Specificity
D007527:Isoenzymes; D046911:Macromolecular Substances; D002262:Carboxy-Lyases; C019972:sulfoalanine decarboxylase