Hepatic synthesis of canavaninosuccinate from ureidohomoserine and aspartate, and its conversion to guanidinosuccinate.
This study continues the exploration of the mechanism for the formation of guanidinoacetate and guanidinosuccinate in the human [Clin. Chem. 21, 235 (1975)]. In this report we describe the formation of canavaninosuccinate from ureidohomoserine and aspartate by a human or bovine liver extract that had high argininosuccinate synthetase (EC 6.3.4.5) activity, and the subsequent formation of guanidinosuccinate by reductive cleavage. In the presence of ATP the optimum pH for the synthetic reaction is 8.4. This reaction can be carried out in either a tris(hydroxymethyl)aminomethane or borate buffer. Subsequent addition of dithiothreitol in the presence of Fe2+ resulted in the cleavage of some of the synthesized canavaninosuccinate to form guanidinosuccinate and homoserine. Synthesis of canavaninosuccinate was strongly inhibited by added argininosuccinate, less so by canavaninosuccinate, arginine, canavanine, glycine, or 2,3-dimercaptopropanol. The Km values for the substrates of the synthetic reaction are 3.6 X 10(-4) mol/liter for aspartate, 1.6 X 10(-3) mol/liter for ureidohomoserine, and 2.92 X 10(-5) mol/liter for ATP. These values are higher than those obtained when the synthesis of argininosuccinate was studied, except for ATP, which yielded a lower value. All of the reactions in the proposed mechanism have now been demonstrated except for the synthesis of canaline from aspartate.
D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism; D002172:Canavanine / Q000031:analogs & derivatives* / Q000378:metabolism; D002417:Cattle; D006146:Guanidines / Q000096:biosynthesis*; D006714:Homoserine / Q000031:analogs & derivatives / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008025:Ligases / Q000378:metabolism; D008099:Liver / Q000378:metabolism*; D013386:Succinates / Q000378:metabolism*; D014508:Urea / Q000031:analogs & derivatives / Q000378:metabolism
D006146:Guanidines; D013386:Succinates; D001224:Aspartic Acid; D002172:Canavanine; D006714:Homoserine; D014508:Urea; D008025:Ligases