Keidel S, Rupp E, Szardenings M
Department of Biochemistry, Medizinische Hochschule Hannover, Federal Republic of Germany.
Eur J Biochem. 1992 Mar 15;204(3):1141-8. doi: 10.1111/j.1432-1033.1992.tb16739.x.
The human retinoic acid receptor alpha was expressed in Escherichia coli. The recombinant protein was found to be very unstable in several E. coli strains, probably due to proteolysis. Conditions were established to obtain reasonable amounts of active protein for ligand and DNA binding studies. The recombinant receptor showed the expected DNA binding activities in gel-retardation assays. Ligand binding properties were measured in a charcoal absorption assay. The dissociation constant for highly specific bound retinoic acid was found to be 0.67 nM. The affinity of several synthetic retinoids to the recombinant protein was determined and compared to their biological activity. Some of the values presented here differ significantly from those published earlier for the receptor or its isolated hormone-binding domain.
人视黄酸受体α在大肠杆菌中表达。发现该重组蛋白在几种大肠杆菌菌株中非常不稳定，可能是由于蛋白水解作用。建立了相关条件以获得用于配体和DNA结合研究的适量活性蛋白。重组受体在凝胶阻滞试验中表现出预期的DNA结合活性。通过活性炭吸附试验测定配体结合特性。发现高度特异性结合视黄酸的解离常数为0.67 nM。测定了几种合成类视黄醇对重组蛋白的亲和力，并将其与它们的生物学活性进行比较。此处给出的一些值与先前发表的关于该受体或其分离的激素结合结构域的值有显著差异。
