Diverse sequences are functional at the C-terminus of the E. coli periplasmic chaperone SurA

被引:7
作者
Chai, Qian [1 ]
Ferrell, Brent [1 ]
Zhong, Meng [1 ]
Zhang, Xinyi [1 ]
Ye, Cui [1 ]
Wei, Yinan [1 ]
机构
[1] Univ Kentucky, Dept Chem, Lexington, KY 40506 USA
基金
美国国家科学基金会;
关键词
mutation; periplasmic molecular chaperone; random sequence library; sequence diversity; OUTER-MEMBRANE PROTEINS; ESCHERICHIA-COLI; PDZ DOMAINS; MOLECULAR CHAPERONE; PEPTIDE RECOGNITION; STATIONARY-PHASE; BIOGENESIS; BINDING; SURVIVAL; PORINS;
D O I
10.1093/protein/gzu003
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
SurA is a major periplasmic molecular chaperone in Escherichia coli and has been shown to assist the biogenesis of several outer membrane proteins. The C-terminal fragment of SurA folds into a short beta-strand, which forms a small three-stranded anti-parallel beta-sheet module with the N-terminal beta-hairpin. We found that the length of the C-terminal fragment, rather than its exact amino acid composition, had a big impact on SurA function. To investigate the determinant factor of the C-terminal sequence, we created a library of SurA constructs randomized in the last 10 residues. We screened the library and randomly analyzed 19 constructs that displayed SurA activity. The C-termini of these constructs shared little sequence similarity, except that beta-strand-forming residues were preferentially enriched. Three SurA constructs were expressed and purified for structural characterization. Circular dichroism and fluorescence spectroscopy analyses revealed that their structures were similar to the structure of the wild-type SurA. Our results suggest that for scaffolding purpose proteins may tolerate various sequences provided certain general requirements such as hydrophobicity and secondary structure propensity are satisfied. Furthermore, the sequence tolerance of SurA at the C-terminus indicates that this area is not likely to be involved in substrate binding.
引用
收藏
页码:111 / 116
页数:6
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