2ON8

Gbeta1 stabilization by in vitro evolution and computational design

2ON8 の概要

• エントリーDOI: 10.2210/pdb2on8/pdb
• 関連するPDBエントリー: 1FCC 1GB4 1P7E 1PGA 1PGB 3GB1
• 分子名称: Immunoglobulin G-binding protein G (2 entities in total)
• 機能のキーワード: beta sheet, alpha helix, improved hydrophobic packing of core residues, protein binding
• 由来する生物種: Streptococcus sp.
• 細胞内の位置: Secreted, cell wall ; Peptidoglycan-anchor : P19909
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 6309.00

構造登録者

Max, K.E.A.,Heinemann, U. (登録日: 2007-01-23, 公開日: 2007-12-04, 最終更新日: 2023-08-30)

主引用文献

Wunderlich, M.,Max, K.E.,Roske, Y.,Mueller, U.,Heinemann, U.,Schmid, F.X.
Optimization of the gbeta1 domain by computational design and by in vitro evolution: structural and energetic basis of stabilization.
J.Mol.Biol., 373:775-784, 2007

PubMed Abstract: Computational design and in vitro evolution are major strategies for stabilizing proteins. For the four critical positions 16, 18, 25, and 29 of the B domain of the streptococcal protein G (Gbeta1), they identified the same optimal residues at positions 16 and 25, but not at 18 and 29. Here we analyzed the energetic contributions of the residues from these two approaches by single and double mutant analyses and determined crystal structures for a variant from the calculation (I16/L18/E25/K29) and from the selection (I16/I18/E25/F29). The structural analysis explains the observed differences in stabilization. Residues 16, 18, and 29 line an invagination, which results from a packing defect between the helix and the beta-sheet of Gbeta1. In all stabilized variants, residues with larger side-chains occur at these positions and packing is improved. In the selected variant, packing is better optimized than in the computed variant. Such differences in side-chain packing strongly affect stability but are difficult to evaluate by computation.

PubMed: 17868696
DOI: 10.1016/j.jmb.2007.08.004

実験手法

X-RAY DIFFRACTION (1.35 Å)