Following evolutionary paths to protein-protein interactions with high affinity and selectivity

被引:66
作者
Levin, Kalia Bernath [1 ,2 ]
Dym, Orly [3 ]
Albeck, Shira [3 ]
Magdassi, Shlomo [2 ]
Keeble, Anthony H. [4 ]
Kleanthous, Colin [4 ]
Tawfik, Dan S. [1 ]
机构
[1] Weizmann Inst Sci, Dept Biol Chem, IL-76100 Rehovot, Israel
[2] Hebrew Univ Jerusalem, Casali Inst Appl Chem, IL-91904 Jerusalem, Israel
[3] Weizmann Inst Sci, Israel Struct Prote Ctr, IL-76100 Rehovot, Israel
[4] Univ York, Dept Biol Area 10, York YO10 5DD, N Yorkshire, England
基金
英国生物技术与生命科学研究理事会;
关键词
IMMUNITY PROTEIN; INTERACTION SPECIFICITY; MOLECULAR-MECHANISMS; CRYSTAL-STRUCTURE; DUAL RECOGNITION; COLICIN; EVOLVABILITY; STABILITY; COMPLEXES; IM9;
D O I
10.1038/nsmb.1670
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
How do intricate multi-residue features such as protein-protein interfaces evolve? To address this question, we evolved a new colicin-immunity binding interaction. We started with Im9, which inhibits its cognate DNase ColE9 at 10(-14) M affinity, and evolved it toward ColE7, which it inhibits promiscuously (K-d > 10(-8) M). Iterative rounds of random mutagenesis and selection toward higher affinity for ColE7, and selectivity ( against ColE9 inhibition), led to an similar to 10(5)-fold increase in affinity and a 10(8)-fold increase in selectivity. Analysis of intermediates along the evolved variants revealed that changes in the binding configuration of the Im protein uncovered a latent set of interactions, thus providing the key to the rapid divergence of new Im7 variants. Overall, protein-protein interfaces seem to share the evolvability features of enzymes, that is, the exploitation of promiscuous interactions and alternative binding configurations via 'generalist' intermediates, and the key role of compensatory stabilizing mutations in facilitating the divergence of new functions.
引用
收藏
页码:1049 / U67
页数:8
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