Chen Liu, Lai Chaohua, Lai Juanhong, Tian Changlin
Hefei National Laboratory for Physical Science at Microscale and School of Life Science, University of Science and Technology of China, Hefei, Anhui, PR China.
Protein Expr Purif. 2011 Apr;76(2):205-10. doi: 10.1016/j.pep.2010.11.007. Epub 2010 Nov 16.
MiRP1 (MinK related protein 1) is a membrane protein in the KCNE family. It can associate with and modulate various voltage gated potassium channels. Mutations in human MiRP1 have been found to cause many congenital and acquired long QT syndromes, which are potentially life-threatening cardiac arrhythmias. Here, human MiRP1 was over-expressed in Escherichia coli, purified and eluted into different detergents. Two dimensional (1)H-(15)N correlated solution nuclear magnetic resonance (NMR) spectra of the human MiRP1 in four different detergent micelles indicated that high resolution solution NMR spectrum can be obtained for human MiRP1 in detergent lyso-myristoylphosphatidylglycerol (LMPG). Circular dichroism (CD) spectroscopy of human MiRP1 indicated a high content of alpha-helical secondary structure in LMPG. Backbone assignments of most MiRP1 residues were achieved through a series of triple resonance NMR experiments. Secondary structure analysis based on backbone chemical shifts showed several stretches of alpha-helices along the primary sequence of MiRP1 in LMPG.
微小相关蛋白1（MiRP1）是KCNE家族中的一种膜蛋白。它可以与多种电压门控钾通道结合并对其进行调节。已发现人类MiRP1中的突变会导致许多先天性和后天性长QT综合征，这些都是潜在的危及生命的心律失常。在此，人类MiRP1在大肠杆菌中过表达，经纯化后洗脱到不同的去污剂中。在四种不同去污剂胶束中的人类MiRP1的二维（1）H-（15）N相关溶液核磁共振（NMR）谱表明，在去污剂溶血肉豆蔻酰磷脂酰甘油（LMPG）中可以获得人类MiRP1的高分辨率溶液NMR谱。人类MiRP1的圆二色性（CD）光谱表明在LMPG中α-螺旋二级结构含量很高。通过一系列三重共振NMR实验实现了大多数MiRP1残基的主链归属。基于主链化学位移的二级结构分析表明，在LMPG中沿着MiRP1的一级序列有几段α-螺旋。
