Interrogating the mechanism of a tight binding inhibitor of AIR carboxylase.
The enzyme aminoimidazole ribonucleotide (AIR) carboxylase catalyzes the synthesis of the purine intermediate, 4-carboxy-5-aminoimidazole ribonucleotide (CAIR). Previously, we have shown that the compound 4-nitro-5-aminoimidazole ribonucleotide (NAIR) is a slow, tight binding inhibitor of the enzyme with a Ki of 0.34 nM. The structural attributes and the slow, tight binding characteristics of NAIR implicated this compound as a transition state or reactive intermediate analog. However, it is unclear what molecular features of NAIR contribute to the mimetic properties for either of the two proposed mechanisms of AIR carboxylase. In order to gain additional information regarding the mechanism for the potent inhibition of AIR carboxylase by NAIR, a series of heterocyclic analogs were prepared and evaluated. We find that all compounds are weaker inhibitors than NAIR and that CAIR analogs are not alternative substrates for the enzyme. Surprisingly, rather subtle changes in the structure of NAIR can lead to profound changes in binding affinity. Computational investigations of enzyme intermediates and these inhibitors reveal that NAIR displays an electrostatic potential surface similar to a proposed reaction intermediate. The result indicates that AIR carboxylase is likely sensitive to the electrostatic surface of reaction intermediates and thus compounds which mimic these surfaces should possess tight binding characteristics. Given the evolutionary relationship between AIR carboxylase and N(5)-CAIR mutase, we believe that this concept extends to the mutase enzyme as well. The implications of this hypothesis for the design of selective inhibitors of the N(5)-CAIR mutase are discussed.
Firestine
Steven M
SM
Department of Pharmaceutical Sciences, Eugene Applebaum, College of Pharmacy and Health Sciences, 259 Mack Avenue, Wayne State University, Detroit, MI 48201, United States. sfirestine@wayne.edu
Wu
Weidong
W
Youn
Hasik
H
Davisson
V Jo
VJ
D000620:Aminoimidazole Carboxamide / Q000031:analogs & derivatives / Q000378:metabolism; D000818:Animals; D002262:Carboxy-Lyases / Q000037:antagonists & inhibitors*; D002645:Chickens; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000494:pharmacology; D006801:Humans; D009593:Nitroimidazoles / Q000138:chemical synthesis / Q000494:pharmacology*; D011485:Protein Binding; D012265:Ribonucleotides / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology*; D055672:Static Electricity
C084132:5-amino-1-(5'-phosphoribofuranosyl)-4-nitroimidazole; D004791:Enzyme Inhibitors; D009593:Nitroimidazoles; D012265:Ribonucleotides; D000620:Aminoimidazole Carboxamide; D002262:Carboxy-Lyases; C022378:phosphoribosylaminoimidazole carboxylase; C031143:AICA ribonucleotide