Partial purification and properties of a transamidinase from Lathyrus sativus seedlings. Involvement in homoarginine metabolism and amine interconversions.
A transamidinase was purified 463-fold from Lathyrus sativus seedlings by affinity chromatography on homoarginine--Sepharose. The enzyme exhibited a wide substrate specificity, and catalysed the reversible transfer of the amidino groups from donors such as arginine, homoarginine and canavanine to acceptors such as lysine, putrescine, agmatine, cadaverine and hydroxylamine. The enzyme could not be detected in the seeds, and attained the highest specific activity in the embryo axis on day 10 after seed germination. Its thiol nature was established by strong inhibition by several thiol blockers and thiol compounds in the presence of ferricyanide. In the absence of an exogenous acceptor, it exhibited weak hydrolytic activity towards arginine. It had apparent mol.wt. 210000, and exhibited Michaelis--Menten kinetics with Km 3.0 mM for arginine. Ornithine competitively inhibited the enzyme, with Ki 1.0 mM in the arginine--hydroxylamine amidino-transfer reaction. Conversion experiments with labelled compounds suggest that the enzyme is involved in homoarginine catabolism during the development of plant embryo to give rise to important amino acids and amine metabolites. Presumptive evidence is also provided for its involvement in the biosynthesis of the guanidino amino acid during seed development. The natural occurrence of arcain in L. sativus and mediation of its synthesis in vitro from agmatine by the transamidinase are demonstrated.
D000217:Acyltransferases / Q000302:isolation & purification; D000579:Amidinotransferases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000588:Amines / Q000378:metabolism*; D000596:Amino Acids / Q000378:metabolism; D019881:Aminoacyltransferases; D001120:Arginine / Q000031:analogs & derivatives*; D002103:Cadaverine / Q000302:isolation & purification; D002846:Chromatography, Affinity; D006709:Homoarginine / Q000378:metabolism*; D010944:Plants / Q000196:embryology / Q000201:enzymology*; D013379:Substrate Specificity
D000588:Amines; D000596:Amino Acids; D006709:Homoarginine; D001120:Arginine; D000579:Amidinotransferases; D000217:Acyltransferases; D019881:Aminoacyltransferases; C023175:transamidases; D002103:Cadaverine