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通过一种新的比色测定法检测硫代次黄苷酸和硫唑嘌呤对5-氨基咪唑-4-甲酰胺核苷酸转甲酰酶的抑制作用。

Detection of inhibition of 5-aminoimidazole-4-carboxamide ribotide transformylase by thioinosinic acid and azathioprine by a new colorimetric assay.

作者信息

Ha T, Morgan S L, Vaughn W H, Eto I, Baggott J E

机构信息

Department of Nutrition Sciences, University of Alabama, Birmingham 35294.

出版信息

Biochem J. 1990 Dec 1;272(2):339-42. doi: 10.1042/bj2720339.

DOI:10.1042/bj2720339
PMID:2268263
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1149705/
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.

摘要

用于5-氨基咪唑-4-甲酰胺核苷酸（AICAR）转甲酰基酶（磷酸核糖氨基咪唑甲酰胺甲酰基转移酶；EC 2.1.2.3）的比色测定法已得到广泛改进。改进后的测定法基于叶酸产物四氢叶酸（H4叶酸）短期被高锰酸盐氧化为对氨基苯甲酰谷氨酸（pABG）。改进后的测定法用于检测外周血单个核细胞（PBMC）粗提物中的转甲酰基酶活性。硫唑嘌呤及其代谢产物硫代次黄嘌呤核苷酸（tIMP）是鸡肝转甲酰基酶和MRL/lpr小鼠（一种系统性自身免疫疾病动物模型）PBMC中转甲酰基酶（相对于AICAR）的竞争性抑制剂。tIMP和硫唑嘌呤对鸡肝酶的Ki值分别为39±4微摩尔和120±10微摩尔，而对MRL/lpr小鼠PBMC中酶的Ki值分别为110±20微摩尔和90±14微摩尔。抗炎药布洛芬和萘普生也是转甲酰基酶的抑制剂。

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Detection of inhibition of 5-aminoimidazole-4-carboxamide ribotide transformylase by thioinosinic acid and azathioprine by a new colorimetric assay.通过一种新的比色测定法检测硫代次黄苷酸和硫唑嘌呤对5-氨基咪唑-4-甲酰胺核苷酸转甲酰酶的抑制作用。
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