Primary structure, chromosomal localization, and functional expression of a voltage-gated sodium channel from human brain.
Primary structure, chromosomal localization, and functional expression of a voltage-gated sodium channel from human brain.
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人脑电压门控钠通道的一级结构、染色体定位和功能表达。
DOI：
10.1073/pnas.89.17.8220
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发表时间：
1992
影响因子：
11.1
通讯作者：
Evans,GA
中科院分区：
文献类型：
--
作者：
Ahmed,CM;Ware,DH;Lee,SC;Patten,CD;Ferrer-Montiel,AV;Schinder,AF;McPherson,JD;Wagner-McPherson,CB;Wasmuth,JJ;Evans,GA
筛选来自人大脑皮层的cDNA文库中钠通道α亚基特异性克隆的存在。三个重叠克隆的连接产生全长cDNA克隆HBA，其提供编码2005个氨基酸的蛋白质的完整核苷酸序列。预测的结构表明四个同源重复序列，并表现出最大的同源性和结构相似性，大鼠脑钠通道II。第二个cDNA克隆，HBB，编码不同亚型的钠通道被分离。来自HBA的3'非翻译区的DNA片段的杂交和使用来自HBB的引物的PCR与人-仓鼠体细胞杂交体将这些克隆定位于人2号染色体。与人类中期染色体的原位杂交将HBA和HBB钠通道的结构基因定位于染色体2 q23 -24.3。通过转染CHO细胞表达钠通道HBA基因产物。所表达的HBA电流具有电压依赖性、钠选择性和河豚毒素敏感性，因此表现出钠通道的生物物理和药理学特性。
A cDNA library derived from human cerebral cortex was screened for the presence of sodium channel alpha subunit-specific clones. Ligation of three overlapping clones generated a full-length cDNA clone, HBA, that provided the complete nucleotide sequence coding for a protein of 2005 amino acids. The predicted structure suggests four homologous repeats and exhibits greatest homology and structural similarity to the rat brain sodium channel II. A second cDNA clone, HBB, that encodes a different subtype of sodium channel was isolated. Hybridization of DNA fragments from the 3' untranslated region of HBA and PCR with primers derived from HBB with human-hamster somatic cell hybrids localized these clones to human chromosome 2. In situ hybridization to human metaphase chromosomes mapped the structural genes for both HBA and HBB sodium channels to chromosome 2q23-24.3. The sodium channel HBA gene product was expressed by transfection in CHO cells. Expressed HBA currents were voltage-dependent, sodium-selective, and tetrodotoxin-sensitive and, thus, exhibit the biophysical and pharmacological properties characteristic of sodium channels.