Imredy J P, MacKinnon R
Howard Hughes Medical Institute, Laboratory of Molecular Neurobiology and Biophysics, New York, NY 10021, USA.
J Mol Biol. 2000 Mar 10;296(5):1283-94. doi: 10.1006/jmbi.2000.3522.
Dendrotoxin proteins isolated from Mamba snake venom block potassium channels with a high degree of specificity and selectivity. Using site-directed mutagenesis we have identified residues that constitute the functional interaction surfaces of delta-dendrotoxin and its voltage-gated potassium channel receptor. delta-Dendrotoxin uses a triangular patch formed by seven side-chains (Lys3, Tyr4, Lys6, Leu7, Pro8, Arg10, Lys26) to block K(+) currents carried by a Shaker potassium channel variant. The inhibitory surface of the toxin interacts with channel residues at Shaker positions 423, 425, 427, 431, and 449 near the pore. Amino acid mutations that interact across the toxin-channel interface were identified by mutant cycle analysis. These results constrain the possible orientation of dendrotoxin with respect to the K(+) channel structure. We propose that dendrotoxin binds near the pore entryway but does not act as a physical plug.
从曼巴蛇毒液中分离出的树突毒素蛋白能高度特异性和选择性地阻断钾通道。通过定点诱变,我们已确定了构成δ-树突毒素及其电压门控钾通道受体功能相互作用表面的残基。δ-树突毒素利用由七个侧链(赖氨酸3、酪氨酸4、赖氨酸6、亮氨酸7、脯氨酸8、精氨酸10、赖氨酸26)形成的三角形区域来阻断由一种摇蚊钾通道变体携带的K(+)电流。毒素的抑制表面与靠近孔道的摇蚊位置423、425、427、431和449处的通道残基相互作用。通过突变循环分析确定了跨毒素-通道界面相互作用的氨基酸突变。这些结果限制了树突毒素相对于K(+)通道结构的可能取向。我们提出树突毒素在孔道入口附近结合,但不作为物理堵塞物起作用。