从大鼠肝脏中纯化线粒体三碘甲状腺原氨酸（T3）受体。

Purification of the mitochondrial triiodothyronine (T3) receptor from rat liver.

作者信息

Sterling K, Campbell G A, Brenner M A

出版信息

Acta Endocrinol (Copenh). 1984 Mar;105(3):391-7. doi: 10.1530/acta.0.1050391.

DOI:10.1530/acta.0.1050391

Abstract

The thyroid hormone receptor of the inner membrane of rat liver mitochondria was purified by osmotic and freeze-thaw lyses followed by partial purification on Sephadex G-200, and then by affinity chromatography with T3-Sepharose 4B. A single predominant protein band demonstrable on sodium dodecylsulphate (SDS) polyacrylamide gel electrophoresis was present in the first 4 mM NaOH elution peak of affinity chromatography. This was collected from affinity peaks from about 30 rat livers followed by preparative polyacrylamide gel electrophoresis. A single absorbance peak was observed by high pressure liquid chromatography (HPLC). The purified protein was analyzed for binding constants, amino acid composition, and characterized by analytical ultracentrifugation. The association constant (KA) exceeded 10(11) M-1. The sedimentation coefficient (S20,W) was 2.2S, partial specific volume (v) 0.72, frictional coefficient (f/fo)s M 1.68 and the molecular weight was estimated at 28 000. The amino acid composition was obtained.

摘要

通过渗透和冻融裂解，然后在Sephadex G - 200上进行部分纯化，再用T3 - Sepharose 4B进行亲和层析，纯化大鼠肝脏线粒体内膜的甲状腺激素受体。在亲和层析的第一个4 mM NaOH洗脱峰中，十二烷基硫酸钠（SDS）聚丙烯酰胺凝胶电泳上可显示出一条单一的主要蛋白带。这是从约30只大鼠肝脏的亲和峰中收集得到的，随后进行制备性聚丙烯酰胺凝胶电泳。通过高压液相色谱（HPLC）观察到一个单一的吸光度峰。对纯化的蛋白质进行结合常数、氨基酸组成分析，并通过分析超速离心进行表征。缔合常数（KA）超过10(11) M-1。沉降系数（S20,W）为2.2S，偏比容（v）为0.72，摩擦系数（f/fo）s M 1.68，分子量估计为28000。得到了氨基酸组成。