ISOLATION OF NEURONAL VOLTAGE SENSITIVE CALCIUM CHANNELS

描述：(改编自申请人的摘要)长期 本应用的目的是阐明其结构和功能 神经元型电压敏感钙通道的多样性 通道蛋白的分子克隆与表达。的可用性 神经元性VSCC的纯亚型对高度 具有治疗多种疾病潜力的选择性拮抗剂 从癫痫到老年痴呆症的各种神经系统疾病。这个 策略包括分离结合的通道蛋白 通过使用涉及凝集素的既定程序来产生芋螺毒素 层析和亲和层析。纯化的蛋白质，它 已经通过亲和交联和光标记鉴定，将被 经十二烷基硫酸钠凝胶电泳法分离后电洗脱并测序。 基于多肽序列的寡核苷酸探针将用于 巴西水仙(Narcine Brasiliens)电子器官基因文库的筛选 并分离出cdna克隆。

DESCRIPTION: (Adapted from the applicant's abstract) The long term objective of this application is to elucidate the structural and functional diversity of neuronal voltage-sensitive calcium channels (VSCC) by molecular cloning and expression of the channel proteins. Availability of pure subtypes of neuronal VSCC is crucial to the development of highly selective antagonists which have great potential for therapy of various neurological diseases ranging from epilepsy to senile dementia. The strategy involves the isolation of the channel protein which binds conotoxin by the use of established procedures involving lectin chromatography and affinity chromatography. The purified protein, which has been identified by affinity cross-linking and photo-labeling, will be electroeluted after separation by SDS gel electrophoresis and sequenced. Oligonucleotide probes based on the peptide sequences will be used to screen a cDNA library of the electric organ of the ray Narcine brasiliensis and isolate cDNA clones.