Stereoselectivity of interaction of phosphoenolpyruvate analogues with various phosphoenolpyruvate-utilizing enzymes.
Stereoselectivity of interaction of phosphoenolpyruvate analogues with various phosphoenolpyruvate-utilizing enzymes.
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磷酸烯醇丙酮酸类似物与各种磷酸烯醇丙酮酸利用酶相互作用的立体选择性。
DOI：
10.1021/bi00299a012
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发表时间：
1984
期刊：
影响因子：
2.9
通讯作者：
Nowak,T
中科院分区：
文献类型：
--
作者：
Duffy,TH;Nowak,T
托马斯H. Duffy 1 * 和托马斯Nowak*· 8摘要：合成并纯化了卤代磷酸烯醇丙酮酸类似物（Z）-磷酸烯醇-3-氟丙酮酸、（E）-磷酸烯醇-3-氟丙酮酸和（Z）-磷酸烯醇-3-溴丙酮酸。在适用的情况下，通过19 F NMR表征类似物。绝对立体选择性的fluorophosphoenolpyruvate异构体作为底物的酶磷酸烯醇式丙酮酸羧激酶，烯醇化酶，丙酮酸磷酸二激酶进行了观察。Z异构体表现出与这些酶的底物活性，而没有底物活性与E异构体进行测量。这两种异构体都表现出丙酮酸激酶的底物活性，然而，与磷酸烯醇丙酮酸作为底物相比，Vm × lx/Km比值大幅降低。用磷酸烯醇丙酮酸羧激酶、烯醇化酶和丙酮酸激酶测定了这些类似物的金属离子依赖性立体选择性抑制作用。阳离子活化剂似乎影响特异性，从而影响这些酶的催化位点。质子纵向弛豫速率滴定表明，thedissociation常数，K3，从酶锰复合物的氟磷酸烯醇式丙酮酸异构体同意，在大多数情况下，与测得的Kx值和类似物结合类似磷酸烯醇式丙酮酸结合。对于磷酸烯醇式丙酮酸羧激酶，（E）-氟磷酸烯醇式丙酮酸的KtK 3表明E异构体的结合受其他底物存在的影响。卤代衍生物显然经历酶-Mn催化的Michael型加成反应，其中溴取代的类似物比氟类似物分解得快得多。
Thomas H. Duffy1***** and Thomas Nowak*· 8 abstract: The halogenated phosphoenolpyruvate analogues (Z)-phosphoenol-3-fluoropyruvate,(£)-phosphoenol-3-fluoropyruvate, and (Z)-phosphoenol-3-bromopyruvate were synthesized and purified. The analogues were characterized by and by 19F NMR where applicable. Absolute stereo-selectivity of the fluorophosphoenolpyruvate isomers as substrates with the enzymes phosphoenolpyruvate carboxykinase, enolase, and pyruvate phosphate dikinase was observed. The Z isomer exhibited substrate activity with these enzymes while no substrate activity was measured with the E isomer. Both isomers exhibited substrate activity with the enzyme pyruvate kinase, however, with a substantial decrease in the Vm¡ lx/Km ratio compared to phosphoenolpyruvate as the substrate. A metal ion dependent stereoselectivity of inhibition was mea-sured for these analogues with the enzymes phosphoenolpyruvate carboxykinase, enolase, andpyruvate kinase. The cation activator appears to affect the specificity and thus the catalytic site of these enzymes. Proton longitudinal relaxation rate titrations demonstrate that thedissociation constants, K3, of the fluorophosphoenolpyruvate isomers from the enzyme-Mn complex agree, in most cases, with the measured Kx values and analogue binding resembles phosphoenolpyruvate binding. With the enzyme phosphoenolpyruvate carboxykinase, the KtK3 for (E)-fluorophosphoenolpyruvate which suggests that the binding of the E isomer is affected by the presence of the other substrates. The halogenated derivatives apparently undergo an enzyme-Mn catalyzed Michael-type addition re-action with the bromo-substituted analogue decomposing much faster than the fluoro analogues.
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影响因子：
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作者：
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作者：
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影响因子：
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作者：
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磷酸烯醇-3-溴丙酮酸酯。
DOI：
10.1016/s0021-9258(18)33318-0
发表时间：
1982
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
M. O'Leary;E. Diaz
通讯作者：
E. Diaz