Differential kinetic properties of L-2-amino-4-methylthio-cis-but-3-enoic acid, a methionine analog inhibitor of S-adenosylmethionine synthetase.

The L-methionine analog, L-2-amino-4-methylthio-cis-but-3-enoic acid (L-cisAMTB), was examined as a potential inhibitor of the enzyme, S-adenosylmethionine (AdoMet) synthetase. The rational design of L-cisAMTB was based on previously observed potent enzyme inhibitory activity for its closely related structural analog, L-2-amino-4-methoxy-cis-but-3-enoic acid (L-cisAMB). The kinetic behavior of L-cisAMTB was studied using AdoMet synthetase isozymes I and II fractionated from L1210 murine leukemia cells. L-cisAMTB, which was a competitive inhibitor with respect to L-methionine, gave apparent Ki values of 21 and 5.7 microM for isozymes I and II, respectively. These values indicate that L-cisAMTB was slightly less inhibitory than L-cisAMB. L-cisAMTB was also a substrate for the AdoMet synthetase reaction, with respective Km values of 555 and 33 microM for isozymes I and II. In the absence of added inhibitors, the activity of isozyme II, but not isozyme I, was stimulated 2.5-fold by the presence of 10% DMSO. This preferential stimulation of isozyme II and the highly significant difference in Km values of L-cisAMTB for isozymes I and II point to possible physical differences in these tumor isozymes that were not apparent in earlier studies.