Initial-Rate Kinetics of Human NMN-Adenylyltransferases:  Substrate and Metal Ion Specificity, Inhibition by Products and Multisubstrate Analogues, and Isozyme Contributions to NAD<sup>+</sup> Biosynthesis

Sorci, Leonardo; Cimadamore, Flavio; Scotti, Stefania; Petrelli, Riccardo; Cappellacci, Loredana; Franchetti, Palmarisa; Orsomando, Giuseppe; Magni, Giulio

doi:10.1021/bi6023379

Cited by 77 publications

(150 citation statements)

References 47 publications

Supporting

Mentioning

142

Contrasting

Order By: Relevance

“…Such dual specificity is characteristic of eukaryotic enzymes from a distinct (albeit evolutionarily and structurally related) PNAT family (36,37). Indeed, both PNAT activities are physiologically relevant for NAD biosynthesis in human cells, which includes de novo and salvage routes proceeding via NaMN or NMN intermediates (36).…”

Section: Resultsmentioning

confidence: 99%

Genomics-driven Reconstruction of Acinetobacter NAD Metabolism

Sorci

Blaby

Ingeniis

et al. 2010

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Enzymes involved in the last steps of NAD biogenesis, nicotinate mononucleotide adenylyltransferase (NadD) and NAD synthetase (NadE), are conserved and essential in most bacterial species and are established targets for antibacterial drug development. Our genomics-based reconstruction of NAD metabolism in the emerging pathogen Acinetobacter baumannii revealed unique features suggesting an alternative targeting strategy. Indeed, genomes of all analyzed Acinetobacter species do not encode NadD, which is functionally replaced by its distant homolog NadM. We combined bioinformatics with genetic and biochemical techniques to elucidate this and other important features of Acinetobacter NAD metabolism using a model (nonpathogenic) strain Acinetobacter baylyi sp. ADP1. Thus, a comparative kinetic characterization of PncA, PncB, and NadV enzymes allowed us to suggest distinct physiological roles for the two alternative, deamidating and nondeamidating, routes of nicotinamide salvage/recycling. The role of the NiaP transporter in both nicotinate and nicotinamide salvage was confirmed. The nondeamidating route was shown to be transcriptionally regulated by an ADP-ribose-responsive repressor NrtR. The NadM enzyme was shown to possess dual substrate specificity toward both nicotinate and nicotinamide mononucleotide substrates, which is consistent with its essential role in all three routes of NAD biogenesis, de novo synthesis as well as the two salvage pathways. The experimentally confirmed unconditional essentiality of nadM provided support for the choice of the respective enzyme as a drug target. In contrast, nadE, encoding a glutamine-dependent NAD synthetase, proved to be dispensable when the nondeamidating salvage pathway functioned as the only route of NAD biogenesis.Acinetobacter baumannii is an emerging pathogen that belongs to a relatively underexplored branch of ␥-proteobacteria. It can cause severe pneumonia and infections of the urinary tract, bloodstream, and other parts of the body. Some isolates of A. baumannii display resistance to many known antibiotics (1, 2), emphasizing the importance of pursuing new therapeutic targets for drug development. Biogenesis of nicotinamide adenine nucleotide (NAD), an indispensable cofactor involved in a multitude of biochemical transformations in metabolic networks of all species, was recently established as a target pathway for the development of new antibiotics (3-7). Beyond its main function as a redox cofactor, NAD is consumed as a co-substrate by a number of nonredox enzymes such as bacterial DNA ligase and protein deacetylase of the CobB/Sir2 family (8, 9). A degradative consumption of NAD by these and, likely, other (not fully elucidated) enzymes demands continuous replenishing of the NAD pool, providing further rationale for targeting essential enzymes involved in its biogenesis and recycling. Among these enzymes, nicotinate mononucleotide adenylyltransferase (NaMNAT) 4 of the NadD family and NAD synthetase of the NadE family are widely recognized as the most promising...

show abstract

Section: Resultsmentioning

confidence: 99%

Genomics-driven Reconstruction of Acinetobacter NAD Metabolism

Sorci

Blaby

Ingeniis

et al. 2010

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…In vertebrate systems, there are differences in the degrees of NaMN versus NMN specificity of three different gene products. More importantly, the genes are differentially regulated and differentially expressed, and their gene products are not expressed in the same cellular compartments (6,59). Thus, there are many cases in which a single EC number corresponds to two or more genes and cases in which a single gene product corresponds to multiple EC numbers.…”

Section: Comparative Genomics Underscores the Rules And Exceptions Inmentioning

confidence: 99%

Microbial NAD Metabolism: Lessons from Comparative Genomics

Gazzaniga

Stebbins

Chang

et al. 2009

Microbiol Mol Biol Rev

199

217

View full text Add to dashboard Cite

SUMMARY NAD is a coenzyme for redox reactions and a substrate of NAD-consuming enzymes, including ADP-ribose transferases, Sir2-related protein lysine deacetylases, and bacterial DNA ligases. Microorganisms that synthesize NAD from as few as one to as many as five of the six identified biosynthetic precursors have been identified. De novo NAD synthesis from aspartate or tryptophan is neither universal nor strictly aerobic. Salvage NAD synthesis from nicotinamide, nicotinic acid, nicotinamide riboside, and nicotinic acid riboside occurs via modules of different genes. Nicotinamide salvage genes nadV and pncA, found in distinct bacteria, appear to have spread throughout the tree of life via horizontal gene transfer. Biochemical, genetic, and genomic analyses have advanced to the point at which the precursors and pathways utilized by a microorganism can be predicted. Challenges remain in dissecting regulation of pathways.

show abstract

“…NadD occupies a central position in bacterial NAD + biosynthesis and is required for both de novo and salvage routes to generate NAD + [14][15][16]. NadD has been recognized as a promising new target for developing novel antibiotics due to its crucial role in synthesizing NAD + [17][18][19][20].…”

Section: Introductionmentioning

confidence: 99%

Biological Activities of the Essential Oil from Erigeron floribundus

et al. 2016

Self Cite

View full text Add to dashboard Cite

Erigeron floribundus (Asteraceae) is an herbaceous plant widely used in Cameroonian traditional medicine to treat various diseases of microbial and non-microbial origin. In the present study, we evaluated the in vitro biological activities displayed by the essential oil obtained from the aerial parts of E. floribundus, namely the antioxidant, antimicrobial and antiproliferative activities. Moreover, we investigated the inhibitory effects of E. floribundus essential oil on nicotinate mononucleotide adenylyltransferase (NadD), a promising new target for developing novel antibiotics, and Trypanosoma brucei, the protozoan parasite responsible for Human African trypanosomiasis. The essential oil composition was dominated by spathulenol (12.2%), caryophyllene oxide (12.4%) and limonene (8.8%). The E. floribundus oil showed a good activity against Staphylococcus aureus (inhibition zone diameter, IZD of 14 mm, minimum inhibitory concentration, MIC of 512 µg/mL). Interestingly, it inhibited the NadD enzyme from S. aureus (IC 50 of 98 µg/mL), with no effects on mammalian orthologue enzymes. In addition, T. brucei proliferation was inhibited with IC 50 values of 33.5 µg/mL with the essential oil and 5.6 µg/mL with the active component limonene. The essential oil exhibited strong cytotoxicity on HCT 116 colon carcinoma cells with an IC 50 value of 14.89 µg/mL, and remarkable ferric reducing antioxidant power (tocopherol-equivalent antioxidant capacity, TEAC = 411.9 µmol·TE/g).

show abstract

Initial-Rate Kinetics of Human NMN-Adenylyltransferases: Substrate and Metal Ion Specificity, Inhibition by Products and Multisubstrate Analogues, and Isozyme Contributions to NAD⁺ Biosynthesis

Cited by 77 publications

References 47 publications

Genomics-driven Reconstruction of Acinetobacter NAD Metabolism

Genomics-driven Reconstruction of Acinetobacter NAD Metabolism

Microbial NAD Metabolism: Lessons from Comparative Genomics

Biological Activities of the Essential Oil from Erigeron floribundus

Contact Info

Product

Resources

About

Initial-Rate Kinetics of Human NMN-Adenylyltransferases: Substrate and Metal Ion Specificity, Inhibition by Products and Multisubstrate Analogues, and Isozyme Contributions to NAD+ Biosynthesis

Cited by 77 publications

References 47 publications

Genomics-driven Reconstruction of Acinetobacter NAD Metabolism

Genomics-driven Reconstruction of Acinetobacter NAD Metabolism

Microbial NAD Metabolism: Lessons from Comparative Genomics

Biological Activities of the Essential Oil from Erigeron floribundus

Contact Info

Product

Resources

About

Initial-Rate Kinetics of Human NMN-Adenylyltransferases: Substrate and Metal Ion Specificity, Inhibition by Products and Multisubstrate Analogues, and Isozyme Contributions to NAD⁺ Biosynthesis