England S K, Uebele V N, Shear H, Kodali J, Bennett P B, Tamkun M M
Department of Molecular Physiology, Vanderbilt University of Medicine, Nashville, TN 37232, USA.
Proc Natl Acad Sci U S A. 1995 Jul 3;92(14):6309-13. doi: 10.1073/pnas.92.14.6309.
Voltage-gated K+ channels are important modulators of the cardiac action potential. However, the correlation of endogenous myocyte currents with K+ channels cloned from human heart is complicated by the possibility that heterotetrameric alpha-subunit combinations and function-altering beta subunits exist in native tissue. Therefore, a variety of subunit interactions may generate cardiac K+ channel diversity. We report here the cloning of a voltage-gated K+ channel beta subunit, hKv beta 3, from adult human left ventricle that shows 84% and 74% amino acid sequence identity with the previously cloned rat Kv beta 1 and Kv beta 2 subunits, respectively. Together these three Kv beta subunits share > 82% identity in the carboxyl-terminal 329 aa and show low identity in the amino-terminal 79 aa. RNA analysis indicated that hKv beta 3 message is 2-fold more abundant in human ventricle than in atrium and is expressed in both healthy and diseased human hearts. Coinjection of hKv beta 3 with a human cardiac delayed rectifier, hKv1.5, in Xenopus oocytes increased inactivation, induced an 18-mV hyperpolarizing shift in the activation curve, and slowed deactivation (tau = 8.0 msec vs. 35.4 msec at -50 mV). hKv beta 3 was localized to human chromosome 3 by using a human/rodent cell hybrid mapping panel. These data confirm the presence of functionally important K+ channel beta subunits in human heart and indicate that beta-subunit composition must be accounted for when comparing cloned channels with endogenous cardiac currents.
电压门控钾通道是心脏动作电位的重要调节因子。然而,由于天然组织中可能存在异源四聚体α亚基组合以及功能改变的β亚基,内源性心肌细胞电流与从人心脏克隆的钾通道之间的相关性变得复杂。因此,多种亚基相互作用可能产生心脏钾通道的多样性。我们在此报告从成人左心室克隆出一种电压门控钾通道β亚基hKvβ3,它与先前克隆的大鼠Kvβ1和Kvβ2亚基的氨基酸序列同一性分别为84%和74%。这三个Kvβ亚基在羧基末端的329个氨基酸中共有>82%的同一性,而在氨基末端的79个氨基酸中同一性较低。RNA分析表明,hKvβ3在人心室中的表达量比心房高2倍,且在健康和患病的人心脏中均有表达。在非洲爪蟾卵母细胞中,将hKvβ3与人心脏延迟整流钾通道hKv1.5共同注射,可增加失活,使激活曲线发生18 mV的超极化偏移,并减慢失活速度(在-50 mV时,τ = 8.0毫秒对35.4毫秒)。通过使用人/啮齿动物细胞杂交定位板,将hKvβ3定位到人类染色体3上。这些数据证实了人心脏中存在功能重要的钾通道β亚基,并表明在将克隆通道与内源性心脏电流进行比较时,必须考虑β亚基的组成。