Sheer D G, Cahnmann H J, Nikodem V M
Biochim Biophys Acta. 1987 Aug 19;930(1):101-6. doi: 10.1016/0167-4889(87)90161-3.
Among the previously reported putative nuclear thyroid hormone receptor forms having molecular masses of 56-59 kDa and 45-49 kDa, respectively, only the former can be the endogenous receptor. The latter must be a degradation product because it is virtually absent in rat liver nuclear extracts prepared in the presence of 20% glycerol and 5 mM Mg2+, which inhibit degradation. In the absence of glycerol, the receptor form of lower mass was present in large amounts in nuclear extracts. Sucrose could not replace glycerol as a protective agent, even in the presence of Mg2+. Thus, the endogenous nuclear thyroid hormone receptor appears to be labile under the experimental conditions used in preparing nuclear extracts. The molecular mass of the nuclear receptor was determined to be 57 kDa on the basis of SDS-polyacrylamide gel electrophoresis after photoaffinity labeling of nuclear proteins with (3,5-125I)-labeled thyroxine.
在先前报道的假定核甲状腺激素受体形式中,分子质量分别为56 - 59 kDa和45 - 49 kDa,只有前者可能是内源性受体。后者必定是一种降解产物,因为在含有20%甘油和5 mM Mg2+(可抑制降解)的条件下制备的大鼠肝核提取物中几乎不存在。在没有甘油的情况下,较低质量的受体形式大量存在于核提取物中。即使存在Mg2+,蔗糖也不能替代甘油作为保护剂。因此,在所使用的制备核提取物的实验条件下,内源性核甲状腺激素受体似乎不稳定。在用(3,5 - 125I)标记的甲状腺素对核蛋白进行光亲和标记后,基于SDS - 聚丙烯酰胺凝胶电泳确定核受体的分子质量为57 kDa。