Experimental and computational studies indicate the mutation of Glu12 to increase the thermostability of oligomeric protease from Pyrococcus horikoshii.

Dongling Zhan, Weiwei Han, Yan Feng

Journal: Journal of molecular modeling 2011;17(6):1241-9

PMID: 20711794

Abstract

The intracellular protease from Pyrococcus horikoshii (PhpI) is a member of the DJ-1/ThiJ/PfpI superfamily, which is suggested to be involved in cellular protection against environmental stresses. In this study, flexible docking approach was employed to dock the ligand into the active site of PhpI. By analyzing the results, active site architecture and certain key residues responsible for substrate specificity were identified on the enzyme. Our docking result indicates that Glu12 plays an important role in substrate binding. The kinetic experiment conducted by Zhan shows that the E12T mutant is more stable than that of the wild-type. We also predict that Glu15, Lys43, and Tyr46 may be important in the catalytic efficiency and thermostability of enzyme. The new structural and mechanistic insights obtained from computational study should be valuable for detailed structures and mechanisms of the member of the DJ-1 superfamily.

Address: Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, Jilin University, Changchun, China.

DOI: 10.1007/s00894-010-0819-0

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https://dx.doi.org/10.1007/s00894-010-0819-0

MeSH Terms: Amino Acid Sequence, Amino Acid Substitution, Bacterial Proteins, Catalytic Domain, Conserved Sequence, Enzyme Stability, Glutamic Acid, Hot Temperature, Models, Molecular, Molecular Sequence Data, Multienzyme Complexes, Mutagenesis, Site-Directed, Peptide Hydrolases, Protein Binding, Protein Structure, Secondary, Protein Structure, Tertiary, Pyrococcus horikoshii, Sequence Alignment