skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: IDENTIFICATION OF NICOTINAMIDE MONONUCLEOTIDE DEAMIDASE OF THE BACTERIAL PYRIDINE NUCLEOTIDE CYCLE REVEALS A NOVEL BROADLY CONSERVED AMIDOHYDROLASE FAMILY

Abstract

The pyridine nucleotide cycle (PNC) is a network of salvage and recycling routes maintaining homeostasis of NAD(P) cofactor pool in the cell. Nicotinamide mononucleotide (NMN) deamidase (EC 3.5.1.42), one of the key enzymes of the bacterial PNC was originally described in Enterobacteria, but the corresponding gene eluded identification for over 30 years. A genomics-based reconstruction of NAD metabolism across hundreds bacterial species suggested that NMN deamidase reaction is the only possible way of nicotinamide salvage in the marine bacterium Shewanella oneidensis. This prediction was verified via purification of native NMN deamidase from S. oneidensis followed by the identification of the respective gene, termed pncC. Enzymatic characterization of the PncC protein, as well as phenotype analysis of deletion mutants, confirmed its proposed biochemical and physiological function in S. oneidensis. Of the three PncC homologs present in E. coli, NMN deamidase activity was confirmed only for the recombinant purified product of the ygaD gene. A comparative analysis at the level of sequence and three dimensional structure, which is available for one of the PncC family member, shows no homology with any previously described amidohydrolases. Multiple alignment analysis of functional and non functional PncC homologs, together with NMN docking experiments, allowed us tomore » tentatively identify the active site area and conserved residues therein. An observed broad phylogenomic distribution of predicted functional PncCs in bacterial kingdom is consistent with a possible role in detoxification of NMN, resulting from NAD utilization by DNA ligase.« less

Authors:
; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1030449
Report Number(s):
PNNL-SA-81056
Journal ID: ISSN 0021-9258; JBCHA3; KP1601030; TRN: US201124%%210
DOE Contract Number:  
AC05-76RL01830
Resource Type:
Journal Article
Journal Name:
Journal of Biological Chemistry
Additional Journal Information:
Journal Volume: 286; Journal Issue: 46; Journal ID: ISSN 0021-9258
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; ALIGNMENT; DETOXIFICATION; DISTRIBUTION; DNA; ENZYMES; FORECASTING; FUNCTIONALS; GENES; HOMEOSTASIS; METABOLISM; MUTANTS; NICOTINAMIDE; NUCLEOTIDES; PHENOTYPE; PURIFICATION; PYRIDINE; RECYCLING; RESIDUES

Citation Formats

Galeazzi, Luca, Bocci, Paolo, Amici, Adolfo, Brunetti, Lucia, Ruggieri, Silverio, Romine, Margaret F., Reed, Samantha B., Osterman, Andrei, Rodionov, Dmitry A., Sorci, Leonardo, and Raffaelli, Nadia. IDENTIFICATION OF NICOTINAMIDE MONONUCLEOTIDE DEAMIDASE OF THE BACTERIAL PYRIDINE NUCLEOTIDE CYCLE REVEALS A NOVEL BROADLY CONSERVED AMIDOHYDROLASE FAMILY. United States: N. p., 2011. Web. doi:10.1074/jbc.M111.275818.
Galeazzi, Luca, Bocci, Paolo, Amici, Adolfo, Brunetti, Lucia, Ruggieri, Silverio, Romine, Margaret F., Reed, Samantha B., Osterman, Andrei, Rodionov, Dmitry A., Sorci, Leonardo, & Raffaelli, Nadia. IDENTIFICATION OF NICOTINAMIDE MONONUCLEOTIDE DEAMIDASE OF THE BACTERIAL PYRIDINE NUCLEOTIDE CYCLE REVEALS A NOVEL BROADLY CONSERVED AMIDOHYDROLASE FAMILY. United States. doi:10.1074/jbc.M111.275818.
Galeazzi, Luca, Bocci, Paolo, Amici, Adolfo, Brunetti, Lucia, Ruggieri, Silverio, Romine, Margaret F., Reed, Samantha B., Osterman, Andrei, Rodionov, Dmitry A., Sorci, Leonardo, and Raffaelli, Nadia. Tue . "IDENTIFICATION OF NICOTINAMIDE MONONUCLEOTIDE DEAMIDASE OF THE BACTERIAL PYRIDINE NUCLEOTIDE CYCLE REVEALS A NOVEL BROADLY CONSERVED AMIDOHYDROLASE FAMILY". United States. doi:10.1074/jbc.M111.275818.
@article{osti_1030449,
title = {IDENTIFICATION OF NICOTINAMIDE MONONUCLEOTIDE DEAMIDASE OF THE BACTERIAL PYRIDINE NUCLEOTIDE CYCLE REVEALS A NOVEL BROADLY CONSERVED AMIDOHYDROLASE FAMILY},
author = {Galeazzi, Luca and Bocci, Paolo and Amici, Adolfo and Brunetti, Lucia and Ruggieri, Silverio and Romine, Margaret F. and Reed, Samantha B. and Osterman, Andrei and Rodionov, Dmitry A. and Sorci, Leonardo and Raffaelli, Nadia},
abstractNote = {The pyridine nucleotide cycle (PNC) is a network of salvage and recycling routes maintaining homeostasis of NAD(P) cofactor pool in the cell. Nicotinamide mononucleotide (NMN) deamidase (EC 3.5.1.42), one of the key enzymes of the bacterial PNC was originally described in Enterobacteria, but the corresponding gene eluded identification for over 30 years. A genomics-based reconstruction of NAD metabolism across hundreds bacterial species suggested that NMN deamidase reaction is the only possible way of nicotinamide salvage in the marine bacterium Shewanella oneidensis. This prediction was verified via purification of native NMN deamidase from S. oneidensis followed by the identification of the respective gene, termed pncC. Enzymatic characterization of the PncC protein, as well as phenotype analysis of deletion mutants, confirmed its proposed biochemical and physiological function in S. oneidensis. Of the three PncC homologs present in E. coli, NMN deamidase activity was confirmed only for the recombinant purified product of the ygaD gene. A comparative analysis at the level of sequence and three dimensional structure, which is available for one of the PncC family member, shows no homology with any previously described amidohydrolases. Multiple alignment analysis of functional and non functional PncC homologs, together with NMN docking experiments, allowed us to tentatively identify the active site area and conserved residues therein. An observed broad phylogenomic distribution of predicted functional PncCs in bacterial kingdom is consistent with a possible role in detoxification of NMN, resulting from NAD utilization by DNA ligase.},
doi = {10.1074/jbc.M111.275818},
journal = {Journal of Biological Chemistry},
issn = {0021-9258},
number = 46,
volume = 286,
place = {United States},
year = {2011},
month = {9}
}