3GJN

Following evolutionary paths to high affinity and selectivity protein-protein interactions using Colicin7 and Immunity proteins

3GJN の概要

• エントリーDOI: 10.2210/pdb3gjn/pdb
• 分子名称: Colicin-E9 immunity protein, Colicin-E7, ZINC ION, ... (4 entities in total)
• 機能のキーワード: protein-protein complex, bacteriocin immunity, antibiotic, antimicrobial, bacteriocin, endonuclease, hydrolase, metal-binding, nuclease
• 由来する生物種: Escherichia coli; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 51880.75

構造登録者

Dym, O.,Tawfik, D.S. (登録日: 2009-03-09, 公開日: 2009-09-15, 最終更新日: 2023-11-01)

主引用文献

Levin, K.B.,Dym, O.,Albeck, S.,Magdassi, S.,Keeble, A.H.,Kleanthous, C.,Tawfik, D.S.
Following evolutionary paths to protein-protein interactions with high affinity and selectivity
Nat.Struct.Mol.Biol., 16:1049-1055, 2009

PubMed Abstract: 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 (Kd > 10(-8) M). Iterative rounds of random mutagenesis and selection toward higher affinity for ColE7, and selectivity (against ColE9 inhibition), led to an approximately 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.

PubMed: 19749752
DOI: 10.1038/nsmb.1670

実験手法

X-RAY DIFFRACTION (2.48 Å)