Turkewitz A P, Amatruda J F, Borhani D, Harrison S C, Schwartz A L
Department of Biochemistry, Harvard University, Cambridge, Massachusetts 02138.
J Biol Chem. 1988 Jun 15;263(17):8318-25.
Human transferrin receptor is a disulfide-linked homodimer of 90-kDa glycoprotein subunits, capable of binding two transferrins. We report a new high yield affinity purification protocol for transferrin receptor from placenta which produces 3-4 mg of highly purified protein. Trypsin cleaves the protein at arginine-121, producing a stable fragment that contains 95% of the extracytoplasmic sequence; similar fragments are produced by several other proteases. The tryptic fragment is a nondisulfide-linked dimer in solution and binds two transferrin molecules. The dimensions of both the dimer fragment and its complex with transferrin are estimated by gel filtration.
人转铁蛋白受体是一种由90 kDa糖蛋白亚基通过二硫键连接而成的同型二聚体,能够结合两个转铁蛋白。我们报告了一种从胎盘中亲和纯化转铁蛋白受体的新的高产方法,该方法可产生3 - 4毫克高度纯化的蛋白。胰蛋白酶在精氨酸-121处切割该蛋白,产生一个稳定的片段,该片段包含95%的胞外序列;其他几种蛋白酶也能产生类似的片段。胰蛋白酶切割片段在溶液中是一种非二硫键连接的二聚体,能结合两个转铁蛋白分子。通过凝胶过滤估算二聚体片段及其与转铁蛋白复合物的尺寸。