Role of side-chains in the cooperative β-hairpin folding of the short C-terminal fragment derived from streptococcal protein G

被引:91
作者
Kobayashi, N
Honda, S
Yoshii, H
Munekata, E [1 ]
机构
[1] Univ Tsukuba, Inst Appl Biochem, Tsukuba, Ibaraki 3058572, Japan
[2] Natl Inst Biosci & Human Technol, Tsukuba, Ibaraki 3058566, Japan
关键词
D O I
10.1021/bi000013p
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
A Short C-terminal fragment of immunoglobulin-binding domain of stroptococcal protein G is known to form nativelike beta-hairpin at physiological conditions. To understand the cooperative folding of the short peptide, eight Ala-substituted mutants of the fragment were investigated with respect to their structural stabilities by analyzing temperature dependence of NMR signals. On comparison of the obtained thermodynamic parameters, we found that the nonpolar residues Tyr45 and Phe52 and the polar residues Asp46 and Thr49 are crucial for the beta-hairpin folding. The results suggest a strong interaction between the nonpolar side chains that participates in a putative hydrophobic cluster and that the polar side chains form a fairly rigid conformation around the loop (46-51). We also investigated the complex formation of the mutants with N-terminal fragment at the variety of temperature to get their thermal unfolding profiles and found that the mutations on the residues Asp36 and Thr39 largely destabilized the complexes, while substitution of Asp47 slightly stabilized the complex. From these results, we deduced that both the hydrophobic cluster formation and the rigidity of the loop (46-51) cooperatively stabilize the beta-hairpin structure of the fragment. These interactions which form a stable beta-hairpin may be the initial structural scaffold which is important in the early folding events of the whole domain.
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页码:6564 / 6571
页数:8
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