Ludwig A, Zong X, Stieber J, Hullin R, Hofmann F, Biel M
Institut für Pharmakologie und Toxikologie der Technischen Universität München, Biedersteiner Strasse 29, 80802 München, Germany.
EMBO J. 1999 May 4;18(9):2323-9. doi: 10.1093/emboj/18.9.2323.
Cardiac pacemaking is produced by the slow diastolic depolarization phase of the action potential. The hyperpolarization-activated cation current (If) forms an important part of the pacemaker depolarization and consists of two kinetic components (fast and slow). Recently, three full-length cDNAs encoding hyperpolarization-activated and cyclic nucleotide-gated cation channels (HCN1-3) have been cloned from mouse brain. To elucidate the molecular identity of cardiac pacemaker channels, we screened a human heart cDNA library using a highly conserved neuronal HCN channel segment and identified two cDNAs encoding HCN channels. The hHCN2 cDNA codes for a protein of 889 amino acids. The HCN2 gene is localized on human chromosome 19p13.3 and contains eight exons spanning approximately 27 kb. The second cDNA, designated hHCN4, codes for a protein of 1203 amino acids. Northern blot and PCR analyses showed that both hHCN2 and hHCN4 are expressed in heart ventricle and atrium. When expressed in HEK 293 cells, either cDNA gives rise to hyperpolarization-activated cation currents with the hallmark features of native If. hHCN2 and hHCN4 currents differ profoundly from each other in their activation kinetics, being fast and slow, respectively. We thus conclude that hHCN2 and hHCN4 may underlie the fast and slow component of cardiac If, respectively.
心脏起搏是由动作电位的缓慢舒张期去极化阶段产生的。超极化激活的阳离子电流(If)构成了起搏去极化的重要部分,由两个动力学成分(快和慢)组成。最近,从小鼠脑中克隆出了三个编码超极化激活且环核苷酸门控阳离子通道(HCN1 - 3)的全长cDNA。为了阐明心脏起搏通道的分子特性,我们使用高度保守的神经元HCN通道片段筛选了人类心脏cDNA文库,并鉴定出两个编码HCN通道的cDNA。hHCN2 cDNA编码一个含有889个氨基酸的蛋白质。HCN2基因定位于人类染色体19p13.3,包含八个外显子,跨度约为27 kb。第二个cDNA,命名为hHCN4,编码一个含有1203个氨基酸的蛋白质。Northern印迹和PCR分析表明,hHCN2和hHCN4在心室和心房中均有表达。当在HEK 293细胞中表达时,任一cDNA都会产生具有天然If特征的超极化激活阳离子电流。hHCN2和hHCN4电流在激活动力学上有很大差异,分别为快和慢。因此我们得出结论,hHCN2和hHCN4可能分别是心脏If快成分和慢成分的基础。