1. Characterization of bacterial NMN deamidase as a Ser/Lys hydrolase expands diversity of serine amidohydrolases.
- Author
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Sorci L, Brunetti L, Cialabrini L, Mazzola F, Kazanov MD, D'Auria S, Ruggieri S, and Raffaelli N
- Subjects
- Amidohydrolases genetics, Amino Acid Sequence, Amino Acid Substitution, Apoenzymes chemistry, Apoenzymes genetics, Catalytic Domain, Conserved Sequence, Enzyme Stability, Escherichia coli Proteins genetics, Kinetics, Models, Molecular, Molecular Sequence Data, Mutagenesis, Site-Directed, Nicotinamide Mononucleotide chemistry, Protein Structure, Secondary, Sequence Homology, Amino Acid, Amidohydrolases chemistry, Escherichia coli Proteins chemistry
- Abstract
NMN deamidase (PncC) is a bacterial enzyme involved in NAD biosynthesis. We have previously demonstrated that PncC is structurally distinct from other known amidohydrolases. Here, we extended PncC characterization by mutating all potential catalytic residues and assessing their individual roles in catalysis through kinetic analyses. Inspection of these residues' spatial arrangement in the active site, allowed us to conclude that PncC is a serine-amidohydrolase, employing a Ser/Lys dyad for catalysis. Analysis of the PncC structure in complex with a modeled NMN substrate supported our conclusion, and enabled us to propose the catalytic mechanism., (Copyright © 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.)
- Published
- 2014
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