Affinity labelling of rat liver acetyl-CoA carboxylase by a 2',3'-dialdehyde derivative of ATP.
The interaction of rat liver acetyl-CoA carboxylase with a 2',3'-dialdehyde derivative of ATP (oATP) has been studied. The degree of the enzyme inactivation has been found to depend on the oATP concentration and the incubation time. ATP was proved to be the only substrate which protected the inactivation. Acetyl-CoA did not effect inactivation, while HCO3- accelerated the process. Ki values for oATP in the absence and presence of HCO3- were 0.35 +/- 0.04 and 0.5 +/- 0.06 mM, and those of the modification constant (kmod) were 0.11 and 0.26 min-1 respectively. oATP completely inhibited the [14C]ADP in equilibrium ATP exchange and did not effect the [14C]acetyl-CoA in equilibrium malonyl-CoA exchange. Incorporation of approximately 1 equivalent of [3H]oATP per acetyl-CoA carboxylase subunit has been shown. No recovery of the modified enzyme activity has been observed in Tris or beta-mercaptoethanol containing buffers, and treatment with NaB3H4 has not led to 3H incorporation. The modification elimination of the ATP triphosphate chain. The results indicated the affinity modification of acetyl-CoA carboxylase by oATP. It was shown that the reagent apparently interacted selectively with the epsilon-amino group of lysine in the ATP-binding site to form a morpholine-like structure.
Rabinkov
A G
AG
V.A. Engelhardt Institute of Molecular Biology, U.S.S.R. Academy of Sciences, Moscow.
Amontov
S V
SV
D000103:Acetyl-CoA Carboxylase / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D000345:Affinity Labels; D000818:Animals; D001639:Bicarbonates; D001665:Binding Sites; D007700:Kinetics; D008025:Ligases / Q000037:antagonists & inhibitors*; D008099:Liver / Q000201:enzymology*; D008239:Lysine / Q000378:metabolism; D051381:Rats; D012545:Schiff Bases
D000345:Affinity Labels; D001639:Bicarbonates; D012545:Schiff Bases; C017199:2',3'-dialdehyde ATP; D000255:Adenosine Triphosphate; D008025:Ligases; D000103:Acetyl-CoA Carboxylase; D008239:Lysine