2W79

Establishing wild-type levels of catalytic activity on natural and artificial (ba)8-barrel protein scaffolds

2W79 の概要

• エントリーDOI: 10.2210/pdb2w79/pdb
• 関連するPDBエントリー: 1QO2 2CFF
• 分子名称: 1-(5-PHOSPHORIBOSYL)-5-[(5-PHOSPHORIBOSYLAMINO) METHYLIDENEAMINO] IMIDAZOLE-4-CARBOXAMIDE ISOMERASE, CHLORIDE ION (3 entities in total)
• 機能のキーワード: isomerase, histidine biosynthesis, amino-acid biosynthesis
• 由来する生物種: THERMOTOGA MARITIMA
• 細胞内の位置: Cytoplasm (By similarity): Q9X0C7
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 54038.02

構造登録者

Claren, J.,Malisi, C.,Hocker, B.,Sterner, R. (登録日: 2008-12-21, 公開日: 2009-02-17, 最終更新日: 2023-12-13)

主引用文献

Claren, J.,Malisi, C.,Hocker, B.,Sterner, R.
Establishing Wild-Type Levels of Catalytic Activity on Natural and Artificial (Beta Alpha)8- Barrel Protein Scaffolds.
Proc.Natl.Acad.Sci.USA, 106:3704-, 2009

PubMed Abstract: The generation of high levels of new catalytic activities on natural and artificial protein scaffolds is a major goal of enzyme engineering. Here, we used random mutagenesis and selection in vivo to establish a sugar isomerisation reaction on both a natural (beta alpha)(8)-barrel enzyme and a catalytically inert chimeric (beta alpha)(8)-barrel scaffold, which was generated by the recombination of 2 (beta alpha)(4)-half barrels. The best evolved variants show turnover numbers and substrate affinities that are similar to those of wild-type enzymes catalyzing the same reaction. The determination of the crystal structure of the most proficient variant allowed us to model the substrate sugar in the novel active site and to elucidate the mechanistic basis of the newly established activity. The results demonstrate that natural and inert artificial protein scaffolds can be converted into highly proficient enzymes in the laboratory, and provide insights into the mechanisms of enzyme evolution.

PubMed: 19237570
DOI: 10.1073/PNAS.0810342106

実験手法

X-RAY DIFFRACTION (1.85 Å)