Hysteretic behavior of methionine adenosyltransferase III. Methionine switches between two conformations of the enzyme with different specific activity.
Methionine adenosyltransferase III (MATIII) catalyzes S-adenosylmethionine (AdoMet) synthesis and, as part of its reaction mechanism, it also hydrolyzes tripolyphosphate. Tripolyphosphatase activity was linear over time and had a slightly sigmoidal behavior with an affinity in the low micromolar range. On the contrary, AdoMet synthetase activity showed a lag phase that was independent of protein concentration but decreased at increasing substrate concentrations. Tripolyphosphatase activity, which appeared to be slower than AdoMet synthesis, was stimulated by preincubation with ATP and methionine so that it matched AdoMet synthetase activity. This stimulation process, which is probably the origin of the lag phase, represents the slow transition between two conformations of the enzyme that could be distinguished by their different tripolyphosphatase activity and sensitivity to S-nitrosylation. Tripolyphosphatase activity appeared to be the rate-determining reaction in AdoMet synthesis and the one inhibited by S-nitrosylation. The methionine concentration necessary to obtain half-maximal stimulation was in the range of physiological methionine fluctuations. Moreover, stimulation of MAT activity by methionine was demonstrated in vivo. We propose that the hysteretic behavior of MATIII, in which methionine induces the transition to a higher specific activity conformation, can be considered as an adaptation to the specific functional requirements of the liver.
del Pino
M M
MM
Division of Hepatology and Gene Therapy, Department of Medicine, University of Navarra, 31008 Pamplona, Navarra, Spain. mspino@unav.es
Corrales
F J
FJ
Mato
J M
JM
D017766:Acid Anhydride Hydrolases / Q000187:drug effects / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D015971:Gene Expression Regulation, Enzymologic; D007527:Isoenzymes / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008297:Male; D008715:Methionine / Q000378:metabolism*; D008716:Methionine Adenosyltransferase / Q000187:drug effects / Q000378:metabolism*; D008956:Models, Chemical; D009569:Nitric Oxide / Q000494:pharmacology; D051381:Rats; D012436:S-Adenosylmethionine / Q000096:biosynthesis*
D007527:Isoenzymes; D009569:Nitric Oxide; D012436:S-Adenosylmethionine; D000255:Adenosine Triphosphate; D008715:Methionine; D008716:Methionine Adenosyltransferase; D017766:Acid Anhydride Hydrolases; C023193:tripolyphosphatase