Binding interface of cardiac potassium channel proteins identified by hydrogen deuterium exchange of synthetic peptides.
Binding interface of cardiac potassium channel proteins identified by hydrogen deuterium exchange of synthetic peptides.
复制标题
通过合成肽的氢氘交换鉴定心脏钾通道蛋白的结合界面。
DOI：
10.1007/s00216-012-5857-2
复制
发表时间：
2012
影响因子：
4.3
通讯作者：
Xiao,Hui
中科院分区：
文献类型：
--
作者：
Chen,Jerri;Angeletti,Ruth;McDonald,ThomasV;Xiao,Hui
通过氢氘交换结合电子转移解离质谱法以单残基分辨率研究了源自人钾通道蛋白 Ether-a-go-go 相关基因 (HERG)、KCNQ1 和 KCNE1 的三种合成肽。 HERG肽前半部分各氨基酸残基掺入氘核的比例高于后半部分，与该肽的核磁共振结构一致，其中氨基酸1-10为柔性螺旋，而氨基酸11-24为稳定的两亲性螺旋。通过比较结合前后序列碎片离子的差异来确定KCNQ1和KCNE1的结合界面。确定参与结合的残基与半胱氨酸交联研究一致，并通过双突变体循环分析证实。
Three synthetic peptides, derived from the human potassium channel proteinsEther-a-go-go-related gene (HERG), KCNQ1, and KCNE1, were investigated by hydrogen deuterium exchange coupled with electron-transfer dissociation mass spectrometry at single residue resolution. Each amino acid residue in the first half of the HERG peptide incorporated deuterons with a higher rate than those in the second half of the peptide, consistent with the nuclear magnetic resonance structure of this peptide, with amino acids 1–10 being a flexible coil, whereas amino acids 11–24 are a stable amphipathic helix. The binding interface of KCNQ1 and KCNE1 was determined by comparing the difference of sequential fragment ions before and after binding. The residues determined to be involved in binding were consistent with a cysteine cross-linking study and confirmed by double mutant cycle analysis.