1EPY

T4 LYSOZYME MUTANT, T21H/C54T/C97A/Q141H/T142H

1EPY の概要

• エントリーDOI: 10.2210/pdb1epy/pdb
• 関連するPDBエントリー: 257L 258L 259L 260L
• 分子名称: LYSOZYME, SULFATE ION, CHLORIDE ION, ... (5 entities in total)
• 機能のキーワード: metal binding, hydrolase
• 由来する生物種: Enterobacteria phage T4
• 細胞内の位置: Host cytoplasm : P00720
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 18902.92

構造登録者

Wray, J.W.,Baase, W.A.,Ostheimer, G.J.,Zhang, X.-J.,Matthews, B.W. (登録日: 2000-03-29, 公開日: 2000-04-12, 最終更新日: 2024-02-07)

主引用文献

Wray, J.W.,Baase, W.A.,Ostheimer, G.J.,Zhang, X.J.,Matthews, B.W.
Use of a non-rigid region in T4 lysozyme to design an adaptable metal-binding site.
Protein Eng., 13:313-321, 2000

PubMed Abstract: It is not easy to find candidate sites within a given protein where the geometry of the polypeptide chain matches that of metal-binding sites in known protein structures. By choosing a location in T4 lysozyme that is inherently flexible, it was possible to engineer a two-histidine site that binds different divalent cations. Crystallographic analysis shows that the geometry of binding of zinc is distorted tetrahedral while that of cobalt and nickel is octahedral. Insofar as spectroscopic data can be measured, they indicate that similar modes of coordination are retained in solution. The two substitutions, Thr21 --> His and Thr142 --> His, lie, respectively, on the surface of the N- and C-terminal domains on opposite sides of the active site cleft. The design takes advantage of hinge-bending motion which allows the binding site to adapt to the most favorable ligand geometry for the metal. Introduction of the two histidines increases the melting temperature of the protein by 2.0 degrees C at pH 7.4. Metal binding further increases the melting temperature, but only by a small amount (up to 1.5 degrees C). A third substitution, Gln141 --> His, which could act as a third ligand in principle, does not do so, demonstrating the difficulty in mimicking naturally occurring metal-binding sites.

PubMed: 10835104
DOI: 10.1093/protein/13.5.313

実験手法

X-RAY DIFFRACTION (1.85 Å)