Knaus H G, McManus O B, Lee S H, Schmalhofer W A, Garcia-Calvo M, Helms L M, Sanchez M, Giangiacomo K, Reuben J P, Smith A B
Department of Membrane Biochemistry and Biophysics, Merck Research Laboratories, Rahway, New Jersey 07065.
Biochemistry. 1994 May 17;33(19):5819-28. doi: 10.1021/bi00185a021.
Tremorgenic indole alkaloids produce neurological disorders (e.g., staggers syndromes) in ruminants. The mode of action of these fungal mycotoxins is not understood but may be related to their known effects on neurotransmitter release. To determine whether these effects could be due to inhibition of K+ channels, the interaction of various indole diterpenes with high-conductance Ca(2+)-activated K+ (maxi-K) channels was examined. Paspalitrem A, paspalitrem C, aflatrem, penitrem A, and paspalinine inhibit binding of [125I]charybdotoxin (ChTX) to maxi-K channels in bovine aortic smooth muscle sarcolemmal membranes. In contrast, three structurally related compounds, paxilline, verruculogen, and paspalicine, enhanced toxin binding. As predicted from the binding studies, covalent incorporation of [125I]ChTX into the 31-kDa subunit of the maxi-K channel was blocked by compounds that inhibit [125I]ChTX binding and enhanced by compounds that stimulate [125I]ChTX binding. Modulation of [125I]ChTX binding was due to allosteric mechanisms. Despite their different effects on binding of [125I]ChTX to maxi-K channels, all compounds potently inhibited maxi-K channels in electrophysiological experiments. Other types of voltage-dependent or Ca(2+)-activated K+ channels examined were not affected. Chemical modifications of paxilline indicate a defined structure-activity relationship for channel inhibition. Paspalicine, a deshydroxy analog of paspalinine lacking tremorgenic activity, also potently blocked maxi-K channels. Taken together, these data suggest that indole diterpenes are the most potent nonpeptidyl inhibitors of maxi-K channels identified to date. Some of their pharmacological properties could be explained by inhibition of maxi-K channels, although tremorgenicity may be unrelated to channel block.
震颤性吲哚生物碱可导致反刍动物出现神经紊乱(如蹒跚综合征)。这些真菌霉菌毒素的作用方式尚不清楚,但可能与其对神经递质释放的已知影响有关。为了确定这些影响是否可能是由于抑制钾离子通道所致,研究了各种吲哚二萜与高电导钙激活钾离子(大电导钾离子通道,maxi-K)通道的相互作用。帕斯帕利特rem A、帕斯帕利特rem C、黄曲霉震颤素、青霉震颤素A和帕斯帕利宁抑制[125I]蝎毒素(ChTX)与牛主动脉平滑肌肌膜中大电导钾离子通道的结合。相比之下,三种结构相关的化合物,即青霉素、疣孢菌素和帕斯帕利辛,增强了毒素结合。正如结合研究所预测的,[125I]ChTX共价掺入大电导钾离子通道的31 kDa亚基被抑制[125I]ChTX结合的化合物所阻断,并被刺激[125I]ChTX结合的化合物所增强。[125I]ChTX结合的调节是由于变构机制。尽管它们对[125I]ChTX与大电导钾离子通道结合的影响不同,但在电生理实验中,所有化合物都能有效抑制大电导钾离子通道。所检测的其他类型的电压依赖性或钙激活钾离子通道未受影响。青霉素的化学修饰表明了通道抑制的明确构效关系。帕斯帕利辛是一种缺乏震颤活性的帕斯帕利宁的脱羟基类似物,也能有效阻断大电导钾离子通道。综上所述,这些数据表明吲哚二萜是迄今为止鉴定出的最有效的非肽类大电导钾离子通道抑制剂。它们的一些药理特性可以通过抑制大电导钾离子通道来解释,尽管震颤性可能与通道阻断无关。