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FEBS Lett. 2014 Mar 18;588(6):1016-23. doi: 10.1016/j.febslet.2014.01.063. Epub 2014 Feb 11.

Characterization of bacterial NMN deamidase as a Ser/Lys hydrolase expands diversity of serine amidohydrolases.

Author information

1
Department of Clinical Sciences, Polytechnic University of Marche, Ancona, Italy.
2
Department of Agricultural, Food and Environmental Sciences, Polytechnic University of Marche, Ancona, Italy.
3
A.A. Kharkevich Institute for Information Transmission Problems, Russian Academy of Sciences, Moscow, Russia.
4
Laboratory for Molecular Sensing, IBP-CNR, Napoli, Italy.
5
Department of Agricultural, Food and Environmental Sciences, Polytechnic University of Marche, Ancona, Italy. Electronic address: n.raffaelli@univpm.it.

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.

KEYWORDS:

Amidohydrolase; Catalytic dyad; NMN deamidase; Pyridine nucleotide; Site-directed mutagenesis

PMID:
24530526
DOI:
10.1016/j.febslet.2014.01.063
[Indexed for MEDLINE]
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