Gerber G E, Anderegg R J, Herlihy W C, Gray C P, Biemann K, Khorana H G
Proc Natl Acad Sci U S A. 1979 Jan;76(1):227-31. doi: 10.1073/pnas.76.1.227.
The sequence of 102 amino acid residues from the NH2 terminus and that of 39 amino acid residues from the COOH terminus of bacteriorhodopsin have been determined. These results are in agreement with those recently published by Ovchinnikov and coworkers [Ovchinnikov, Y.A., Abdulaey, N.G., Feigina, M.Y., Kiselev, A.V. & Lobanov, N.A. (1977) FEBS Lett. 84, 1-4]. Chymotryptic cleavage of bacteriorhodopsin produced two fragments, C-1 (Mr 19,000) and C-2 (Mr 6900), the latter containing the blocked NH2 terminus (pyroglutamic acid). Further fragmentation with CNBr gave mostly hydrophobic fragments, which were separated by gel permeation and reverse-phase high-pressure liquid chromatography in formic acid/ethanol/water mixtures. The fragments were sequenced by a judicious combination of mass spectrometric peptide sequencing and automated Edman degradation. The C-2 fragments were ordered on the basis of methionine-containing peptides identified by gas chromatographic mass spectrometry, while C-1 and C-2 were arranged by analysis of an overlapping CNBr fragment.
已经确定了细菌视紫红质从NH2末端起的102个氨基酸残基序列以及从COOH末端起的39个氨基酸残基序列。这些结果与奥夫钦尼科夫及其同事最近发表的结果一致[奥夫钦尼科夫，Y.A.，阿卜杜拉耶夫，N.G.，费吉娜，M.Y.，基谢廖夫，A.V. & 洛巴诺夫，N.A.（1977年）《欧洲生物化学学会联合会快报》84，1 - 4]。细菌视紫红质经胰凝乳蛋白酶裂解产生两个片段，C - 1（分子量19,000）和C - 2（分子量6900），后者含有被封闭的NH2末端（焦谷氨酸）。用溴化氰进一步裂解主要产生疏水片段，这些片段通过凝胶渗透和在甲酸/乙醇/水混合物中的反相高压液相色谱法进行分离。通过质谱肽测序和自动埃德曼降解的明智组合对这些片段进行测序。基于通过气相色谱质谱法鉴定的含甲硫氨酸肽段对C - 2片段进行排序，而通过分析一个重叠的溴化氰片段对C - 1和C - 2进行排列。
