title: Detection of inhibition of 5-aminoimidazole-4-carboxamide ribotide transformylase by thioinosinic acid and azathioprine by a new colorimetric assay.
abstract: The colorimetric assay for 5-aminoimidazole-4-carboxamide ribotide (AICAR) transformylase (phosphoribosylamino-imidazolecarboxamide formyltransferase; EC 2.1.2.3) has been extensively modified. The modified assay is based upon the short-term permanganate oxidation of the folate product, tetrahydrofolate (H4folate) to p-aminobenzoyl glutamate (pABG). The modified assay was used to detect the transformylase activity in crude extracts of peripheral-blood mononuclear cells (PBMCs). Azathioprine and its metabolite, thioinosinic acid (tIMP), are competitive inhibitors (with respect to AICAR) of the chicken liver transformylase and the transformylase from PBMCs of the MRL/lpr mouse, an animal model of systemic autoimmune disease. The Ki values of tIMP and azathioprine for the chicken liver enzyme are 39 +/- 4 microM and 120 +/- 10 microM, whereas the Ki values for the enzyme from PBMCs of the MRL/lpr mouse are 110 +/- 20 microM and 90 +/- 14 microM respectively. The anti-inflammatory drugs ibuprofen and naproxen are also inhibitors of the transformylase.
authors: Department of Nutrition Sciences, University of Alabama, Birmingham 35294.
authors: T
authors: T
authors: Ha
authors: S L
authors: SL
authors: Morgan
authors: W H
authors: WH
authors: Vaughn
authors: I
authors: I
authors: Eto
authors: J E
authors: JE
authors: Baggott
mesh_terms: D000217:Acyltransferases / Q000037:antagonists & inhibitors* / Q000097:blood; D000818:Animals; D001327:Autoimmune Diseases / Q000097:blood / Q000201:enzymology; D001379:Azathioprine / Q000494:pharmacology*; D001667:Binding, Competitive; D002645:Chickens; D003124:Colorimetry / Q000379:methods; D005260:Female; D006801:Humans; D019877:Hydroxymethyl and Formyl Transferases; D007291:Inosine Monophosphate / Q000031:analogs & derivatives* / Q000494:pharmacology; D007700:Kinetics; D007963:Leukocytes, Mononuclear / Q000201:enzymology*; D008099:Liver / Q000201:enzymology*; D051379:Mice; D008817:Mice, Mutant Strains; D051027:Phosphoribosylaminoimidazolecarboxamide Formyltransferase; D013873:Thionucleotides / Q000494:pharmacology*
chemical_list: D013873:Thionucleotides; D007291:Inosine Monophosphate; C003449:thioinosinic acid; D019877:Hydroxymethyl and Formyl Transferases; D051027:Phosphoribosylaminoimidazolecarboxamide Formyltransferase; D000217:Acyltransferases; D001379:Azathioprine