Spencer S A, Hammonds R G, Henzel W J, Rodriguez H, Waters M J, Wood W I
Department of Developmental Biology, Genentech, Inc., South San Francisco, California 94080.
J Biol Chem. 1988 Jun 5;263(16):7862-7.
A putative growth hormone receptor from detergent-solubilized rabbit liver membranes and the growth hormone binding protein from rabbit serum have been purified 59,000- and 400,000-fold, respectively, primarily by affinity chromatography. Both purified proteins exhibit high affinity binding for human growth hormone; K alpha = 9-30 x 10(9) M-1 for the liver receptor and K alpha = 6 x 10(9) M-1 for the binding protein. The apparent molecular weight of the liver receptor is 130,000 by reduced sodium dodecyl sulfate gel electrophoresis, while that of the binding protein is 51,000. Both contain N-linked carbohydrate. The amino-terminal sequences of the liver growth hormone receptor and the serum binding protein were found to be the same, indicating that the binding protein corresponds to the extracellular domain of the liver receptor. Ubiquitin was found covalently linked to the liver receptor but not to the serum binding protein. The amino acid sequences of several peptides from the liver receptor were also determined after tryptic and V8 protease digestion.
从去污剂溶解的兔肝细胞膜中得到的一种假定生长激素受体和兔血清中的生长激素结合蛋白，分别通过亲和层析法纯化了59,000倍和400,000倍。两种纯化蛋白均对人生长激素表现出高亲和力结合；肝受体的Kα = 9 - 30×10⁹ M⁻¹，结合蛋白的Kα = 6×10⁹ M⁻¹。经还原十二烷基硫酸钠凝胶电泳测定，肝受体的表观分子量为130,000，而结合蛋白的表观分子量为51,000。二者均含有N - 连接的碳水化合物。发现肝生长激素受体和血清结合蛋白的氨基末端序列相同，表明结合蛋白对应于肝受体的细胞外结构域。发现泛素与肝受体共价连接，但与血清结合蛋白不连接。在胰蛋白酶和V8蛋白酶消化后，还测定了肝受体几种肽段的氨基酸序列。
