SYNTHESIS OF A REACTIVE ATP ANALOG AND ITS PRELIMINARY APPLICATION AS AN AFFINITY LABELING REAGENT
SYNTHESIS OF A REACTIVE ATP ANALOG AND ITS PRELIMINARY APPLICATION AS AN AFFINITY LABELING REAGENT
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DOI：
10.1093/oxfordjournals.jbchem.a131947
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发表时间：
1978-01-01
影响因子：
2.7
通讯作者：
IMAHORI, K
中科院分区：
文献类型：
--
作者：
NAGATA, K;SUZUKI, K;IMAHORI, K
反应性ATP类似物，N6-（6-溴乙酰氨基己基）-AMP。PCP [腺苷酰-（β，.伽马亚甲基）-二膦酸酯]，试图共价标记各种酶的腺嘌呤核苷酸，特别是ATP的结合位点，所述酶利用腺嘌呤核苷酸作为底物、辅因子、抑制剂或变构效应物。该试剂在非常温和的条件下快速灭活兔肌甘油醛3-磷酸脱氢酶（GPD）、肌激酶和肌酸激酶。腺嘌呤核苷酸底物阻止失活。在GPD的情况下，当每摩尔酶亚基掺入1摩尔试剂时，观察到完全失活。本发明的ATP类似物可用作各种腺嘌呤核苷酸依赖性酶的亲和标记试剂。
A reactive ATP analog, N6-(6-bromoacetamidohexyl)-AMP .cntdot. PCP [adenylyl-(.beta.,.gamma.-methylene)-diphosphonate], was synthesized in an attempt to covalently label the binding sites for adenine nucleotides, especially ATP, of various enzymes which utilize adenine nucleotides as substrates, cofactors, inhibitors or allosteric effectors. This reagent rapidly inactivated rabbit muscle glyceraldehyde 3-phosphate dehydrogenase (GPD), myokinase and creatine kinase under very mild conditions. Adenine nucleotide substrates prevented the inactivation. In the case of GPD, complete inactivation was observed when 1 mol of the reagent per mol of enzyme subunit was incorporated into the enzyme. The present ATP analog may be useful as an affinity labeling reagent for various adenine nucleotide-dependent enzymes.