Probing the interaction of lipids with the non-annular binding sites of the potassium channel KcsA by magic-angle spinning NMR.
Probing the interaction of lipids with the non-annular binding sites of the potassium channel KcsA by magic-angle spinning NMR.
复制标题
DOI：
10.1016/j.bbamem.2011.09.017
复制
发表时间：
2012-01
影响因子：
3.4
通讯作者：
Williamson, Philip T. F.
中科院分区：
文献类型：
--
作者：
Marius, Phedra;de Planque, Maurits R. R.;Williamson, Philip T. F.
关键词：
钾通道KcsA的活性通过阴离子脂质与位于通道亚基之间的界面处的高亲和力非环状脂质结合位点的相互作用来严格调节。在这里，我们提出了固态磷NMR的研究，解决了带负电荷的脂质磷脂酰甘油内的非环状脂质结合位点。磷脂酰甘油结合后观察到的化学位移扰动表明非环状结合位点内带正电荷的侧链与带负电荷的脂质头基之间的相互作用。定点诱变研究将这些电荷相互作用归因于R64和R89。在功能上，从R64和R89去除正电荷似乎协同作用以降低通道开放的可能性。固态31 P MAS NMR研究已被用于研究与KcsA的非环状结合位点的结合。已鉴定残基R64和R89在非环状结合位点中识别阴离子脂质中是重要的。单通道电流记录证明了这些相互作用对KcsA活性的功能重要性。
The activity of the potassium channel KcsA is tightly regulated through the interactions of anionic lipids with high-affinity non-annular lipid binding sites located at the interface between the channel's subunits. Here we present solid-state phosphorous NMR studies that resolve the negatively charged lipid phosphatidylglycerol within the non-annular lipid-binding site. Perturbations in chemical shift observed upon the binding of phosphatidylglycerol are indicative of the interaction of positively charged sidechains within the non-annular binding site and the negatively charged lipid headgroup. Site directed mutagenesis studies have attributed these charge interactions to R64 and R89. Functionally the removal of the positive charges from R64 and R89 appears to act synergistically to reduce the probability of channel opening. ► Solid-state 31P MAS NMR studies have been used to characterise binding to the non-annular binding site of KcsA. ► Residues R64 and R89 have been identified as important in the recognition of anionic lipids in the non-annular binding site. ► Single-channel current recordings demonstrate the functional importance of these interactions for KcsA activity.
登录
查看更多内容
影响因子：
4.8
作者：
Bonev, BB;Breukink, E;Watts, A
通讯作者：
Watts, A
影响因子：
2.9
作者：
PINHEIRO, TJT;WATTS, A
通讯作者：
WATTS, A
影响因子：
5.6
作者：
Bokvist, M;Lindström, F;Gröbner, G
通讯作者：
Gröbner, G
DOI：
10.1007/s00249-009-0512-3
发表时间：
2010-03
影响因子：
2
作者：
Marsh, Derek
通讯作者：
Marsh, Derek
DOI：
10.1085/jgp.114.4.551
发表时间：
1999-10
期刊：
The Journal of general physiology
影响因子：
--
作者：
Heginbotham L;LeMasurier M;Kolmakova-Partensky L;Miller C
通讯作者：
Miller C