Schenk D B, Phelps M N, Porter J G, Fuller F, Cordell B, Lewicki J A
Proc Natl Acad Sci U S A. 1987 Mar;84(6):1521-5. doi: 10.1073/pnas.84.6.1521.
A receptor for atrial natriuretic peptide (ANP) was purified 2700-fold, to apparent homogeneity, from cultured bovine aortic smooth muscle cells by affinity chromatography. The native ANP receptor has a molecular weight of 125,000 as determined by both metrizamide gradient centrifugation and nonreducing NaDodSO4/polyacrylamide gel electrophoresis. With 125I-labeled ANP as ligand, the purified receptor bound a maximum of 5.70 nmol of ligand per mg of protein and the dissociation constant was 4.0 X 10(-10)M. Upon treatment with 10 mM dithiothreitol, the purified receptor migrated as a single band at Mr 60,500 in NaDodSO4/polyacrylamide gel electrophoresis. These findings show that the holoreceptor for ANP in vascular tissue is composed of two subunits of identical apparent molecular weight, presumably linked by a disulfide bridge(s).
通过亲和层析从培养的牛主动脉平滑肌细胞中纯化出一种心房利钠肽（ANP）受体，纯化倍数达2700倍，达到表观均一性。通过甲泛葡胺梯度离心法和非还原型十二烷基硫酸钠/聚丙烯酰胺凝胶电泳法测定，天然ANP受体的分子量为125,000。以125I标记的ANP作为配体，纯化后的受体每毫克蛋白质最多结合5.70纳摩尔配体，解离常数为4.0×10(-10)M。用10 mM二硫苏糖醇处理后，纯化后的受体在十二烷基硫酸钠/聚丙烯酰胺凝胶电泳中迁移为一条位于Mr 60,500的单带。这些发现表明，血管组织中ANP的全受体由两个表观分子量相同的亚基组成，推测通过二硫键相连。
