CONFORMATION OF PROLINE RESIDUES IN BACTERIORHODOPSIN

被引:17
作者
DEBER, CM [1 ]
SORRELL, BJ [1 ]
XU, GY [1 ]
机构
[1] UNIV TORONTO,DEPT BIOCHEM,TORONTO M5S 1A8,ONTARIO,CANADA
基金
加拿大自然科学与工程研究理事会; 英国医学研究理事会;
关键词
D O I
10.1016/0006-291X(90)90755-C
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Proline, noted as a hydrophilic residue with helix-breaking potential, nevertheless occurs widely in putatively α-helical transmembrane segments of many transport proteins. Ligand-activated or enzyme-assisted trans/cis isomerization of an X-proline peptide bond (where X=any amino acid) - a dynamic, reversible event which could alter the orientation of a transmembrane α-helix - may provide the molecular basis for a protein channel regulatory process. Further elucidation of such a function requires knowledge of the isomeric status of the X-Pro bonds in native conformations of membrane proteins. We have used 13C nuclear magnetic resonance (NMR) spectroscopy to examine the conformation of intramembranous X-Pro peptide bonds in biosynthetically-labelled samples of a model transport protein, bacteriorhodopsin (bR) (purple membrane). Spectra of 13C-Tyr-carbonyl labelled bR (in the solvent system CHCl3:CD3OD (1:1) + 0.1 M LiClO4) first established that all 11 bR Tyr residues were sufficiently mobile for their resonances to be detected and resolved, independent of their domain location within the bR sequence. By taking advantage of the known diagnostic chemical shifts of the isomers of Pro-Cγ carbon resonances, spectra of bR labelled with 13Cγ-Pro were then used to demonstrate that all 11 bR X-Pro peptide bonds - including those within the protein's membrane domain (Pro50, Pro91, Pro186) - are in the trans conformation in resting state bR. © 1990.
引用
收藏
页码:862 / 869
页数:8
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