Wang J K, McDowell J H, Hargrave P A
Biochemistry. 1980 Oct 28;19(22):5111-7. doi: 10.1021/bi00563a027.
A dipeptide containing the binding site for retinal in bovine rhodopsin has been isolated and its sequence determined. Rhodopsin containing [11-3H]retinal was prepared in chromatographically pure form, and the [3H]retinal was reductively linked to its binding site on opsin by using borane--dimethylamine. The [3H]retinylopsin in octyl glucoside was exhaustively digested with Pronase, and its peptides were separated on silica gel in chloroform/methanol/ammonia [Bownds, D. (1967) Nature (London) 216, 1178--1181] followed by silica gel thin-layer chromatography in two solvent systems. The major retinyl peptide was shown to be alanyl-N epsilon-retinyllysine by amino acid composition, 3H content, and amino acid sequence analysis. The retinyl binding site is located in the carboxyl-terminal region of rhodopsin: when rod cell disk membranes containing [3H]retinal rhodopsin were digested with thermolysin and then reacted with sodium borohydride or borane--dimethylamine, [3H]retinal was reduced onto the F2 (Mr congruent to 6000) fragment, which derives from rhodopsin's carboxyl-terminal region.
一种包含牛视紫红质中视黄醛结合位点的二肽已被分离出来并确定了其序列。制备了色谱纯形式的含有[11-³H]视黄醛的视紫红质，通过使用硼烷-二甲胺将[³H]视黄醛还原连接到视蛋白上的结合位点。用链霉蛋白酶将辛基葡糖苷中的[³H]视黄醛视蛋白彻底消化，其肽段在硅胶上于氯仿/甲醇/氨中分离[鲍恩兹，D.（1967年）《自然》（伦敦）216, 1178 - 1181]，随后在两种溶剂系统中进行硅胶薄层层析。通过氨基酸组成、³H含量和氨基酸序列分析表明，主要的视黄醛肽为丙氨酰-Nε-视黄醛赖氨酸。视黄醛结合位点位于视紫红质的羧基末端区域：当用嗜热菌蛋白酶消化含有[³H]视黄醛视紫红质的视杆细胞盘膜，然后与硼氢化钠或硼烷-二甲胺反应时，[³H]视黄醛被还原到源自视紫红质羧基末端区域的F2（分子量约为6000）片段上。
