Schlichter L C, Pahapill P A, Chung I
Department of Physiology, University of Toronto, Ontario.
J Pharmacol Exp Ther. 1992 May;261(2):438-46.
Originally developed as antidotes to organophosphorus nerve poisons, the oximes have attracted renewed interest in studies of cellular regulation. In particular, 2,3-butanedione monoxime (BDM) has gained attention as a useful membrane-permeant "chemical phosphatase" for studying roles of protein phosphorylation. It has been proposed that effects of BDM on cardiac muscle tension, action potentials, neuromuscular transmission and ion currents are related to dephosphorylation of substrates as diverse as myofibrils and ion channels. In the present study, voltage-dependent K+ currents in human T lymphocytes were studied using the whole cell patch clamp technique. Preincubating intact cells briefly in 5 mM BDM before recording reduced the K+ current in an irreversible manner, consistent with chemical (phosphatase?) modification of the channels. In contrast, acute BDM treatment produced a rapid, reversible block of K+ current with half block at about 5 mM. Moreover, including adenosine-O-5'-(3-thiotriphosphate) (500 microM) in the patch pipette did not prevent the rapid, reversible block by BDM. Under these conditions, the most likely mechanism was a direct block of channels from the outside. Because similar K+ currents are present in many tissue and cell types, a direct channel block suggests caution in interpreting the effects of oximes as resulting from protein dephosphorylation.
肟最初是作为有机磷神经毒剂的解毒剂而开发的,现在在细胞调节研究中重新引起了人们的兴趣。特别是,2,3-丁二酮一肟(BDM)作为一种有用的可透过膜的“化学磷酸酶”,在研究蛋白质磷酸化的作用方面受到了关注。有人提出,BDM对心肌张力、动作电位、神经肌肉传递和离子电流的影响与肌原纤维和离子通道等多种底物的去磷酸化有关。在本研究中,使用全细胞膜片钳技术研究了人T淋巴细胞中的电压依赖性钾电流。在记录前将完整细胞在5 mM BDM中短暂预孵育会以不可逆的方式降低钾电流,这与通道的化学(磷酸酶?)修饰一致。相比之下,急性BDM处理会快速、可逆地阻断钾电流,半数阻断浓度约为5 mM。此外,在膜片吸管中加入腺苷-O-5'-(3-硫代三磷酸)(500 microM)并不能阻止BDM的快速、可逆阻断。在这些条件下,最可能的机制是从外部直接阻断通道。由于许多组织和细胞类型中都存在类似的钾电流,因此直接的通道阻断表明在将肟的作用解释为蛋白质去磷酸化的结果时应谨慎。