2W79

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

Summary for 2W79

• Entry DOI: 10.2210/pdb2w79/pdb
• Related: 1QO2 2CFF
• Descriptor: 1-(5-PHOSPHORIBOSYL)-5-[(5-PHOSPHORIBOSYLAMINO) METHYLIDENEAMINO] IMIDAZOLE-4-CARBOXAMIDE ISOMERASE, CHLORIDE ION (3 entities in total)
• Functional Keywords: isomerase, histidine biosynthesis, amino-acid biosynthesis
• Biological source: THERMOTOGA MARITIMA
• Cellular location: Cytoplasm (By similarity): Q9X0C7
• Total number of polymer chains: 2
• Total formula weight: 54038.02

Authors

Claren, J.,Malisi, C.,Hocker, B.,Sterner, R. (deposition date: 2008-12-21, release date: 2009-02-17, Last modification date: 2023-12-13)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (1.85 Å)