Studies of reversible inhibition, irreversible inhibition, and activation of alkaline phosphatase by capillary electrophoresis
Studies of reversible inhibition, irreversible inhibition, and activation of alkaline phosphatase by capillary electrophoresis
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DOI：
10.1016/s0003-2697(02)00062-3
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发表时间：
2002-08-15
影响因子：
2.9
通讯作者：
Gilman, SD
中科院分区：
文献类型：
--
作者：
Whisnant, AR;Gilman, SD
用毛细管电泳法研究了小牛肠碱性磷酸酶（3.1.3.1）的可逆抑制、不可逆抑制和激活。毛细管电泳酶抑制剂分析是基于在底物填充的毛细管中的抑制剂和酶区的电泳混合。通过激光诱导荧光在毛细管中检测到的产物形成减少来指示酶抑制。可逆酶抑制剂可以通过电泳数据的Michaelis-Menten处理来定量。研究了钒酸钠和砷酸钠对碱性磷酸酶的可逆竞争性抑制，以及茶碱对碱性磷酸酶的可逆非竞争性抑制。使用毛细管电泳测定的这些可逆抑制剂的Ki值在文献中报道的相同酶-抑制剂组合的值的范围内。已观察到EDTA在1.0 mM及以上浓度下对碱性磷酸酶的不可逆抑制。还观察到EDTA在20至400 μ M浓度下激活碱性磷酸酶。(C)2002 Elsevier Science（美国）。All rights reserved.
Reversible inhibition, irreversible inhibition, and activation of calf intestinal alkaline phosphatase (EC 3.1.3.1) have been studied by capillary electrophoresis. The capillary electrophoretic enzyme-inhibitor assays were based on electrophoretic mixing of inhibitor and enzyme zones in a substrate-filled capillary. Enzyme inhibition was indicated by a decrease in product formation detected in the capillary by laser-induced fluorescence. Reversible enzyme inhibitors could be quantified by Michaelis-Menten treatment of the electrophoretic data. Reversible, competitive inhibition of alkaline phosphatase by sodium vanadate and sodium arsenate has been examined, and reversible, noncompetitive inhibition by theophylline has been studied. The K-i values determined for these reversible inhibitors using capillary electrophoresis are within the range of values reported in the literature for the same enzyme-inhibitor combinations. Irreversible inhibition of alkaline phosphatase by EDTA at concentrations of 1.0 mM and above has been observed. Activation of alkaline phosphatase has also been observed for EDTA at concentrations from 20 to 400 muM. (C) 2002 Elsevier Science (USA). All rights reserved.