2P05

Structural Insights into the Evolution of a Non-Biological Protein

2P05 の概要

• エントリーDOI: 10.2210/pdb2p05/pdb
• 関連するPDBエントリー: 2P09
• 分子名称: a non-biological ATP binding protein 1819, ZINC ION, CHLORIDE ION, ... (5 entities in total)
• 機能のキーワード: alpha/beta fold, de novo protein
• 由来する生物種: unidentified
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 10489.43

構造登録者

Smith, M.,Rosenow, M.,Wang, M.,Allen, J.P.,Szostak, J.W.,Chaput, J.C. (登録日: 2007-02-28, 公開日: 2007-06-05, 最終更新日: 2024-02-21)

主引用文献

Smith, M.D.,Rosenow, M.A.,Wang, M.,Allen, J.P.,Szostak, J.W.,Chaput, J.C.
Structural insights into the evolution of a non-biological protein: importance of surface residues in protein fold optimization.
PLoS ONE, 2:e467-e467, 2007

PubMed Abstract: Phylogenetic profiling of amino acid substitution patterns in proteins has led many to conclude that most structural information is carried by interior core residues that are solvent inaccessible. This conclusion is based on the observation that buried residues generally tolerate only conserved sequence changes, while surface residues allow more diverse chemical substitutions. This notion is now changing as it has become apparent that both core and surface residues play important roles in protein folding and stability. Unfortunately, the ability to identify specific mutations that will lead to enhanced stability remains a challenging problem. Here we discuss two mutations that emerged from an in vitro selection experiment designed to improve the folding stability of a non-biological ATP binding protein. These mutations alter two solvent accessible residues, and dramatically enhance the expression, solubility, thermal stability, and ligand binding affinity of the protein. The significance of both mutations was investigated individually and together, and the X-ray crystal structures of the parent sequence and double mutant protein were solved to a resolution limit of 2.8 and 1.65 A, respectively. Comparative structural analysis of the evolved protein to proteins found in nature reveals that our non-biological protein evolved certain structural features shared by many thermophilic proteins. This experimental result suggests that protein fold optimization by in vitro selection offers a viable approach to generating stable variants of many naturally occurring proteins whose structures and functions are otherwise difficult to study.

PubMed: 17520026
DOI: 10.1371/journal.pone.0000467

実験手法

X-RAY DIFFRACTION (2.8 Å)