Article Summary:
Title: Identification of Inactive Mutants of NMN Deamidase as Molecular Recognition Elements for an NMN-Specific Biosensor
Introduction:
- Nicotinamide mononucleotide (NMN) is an important intermediate in the production of nicotinamide adenine dinucleotide (NAD+), a key molecule involved in cellular metabolism.
- Supplementation of NMN has shown positive effects on various diseases, such as metabolic, vascular, and neurodegenerative disorders.
Study Objective:
- This study aimed to identify inactive mutants of NMN deamidase (PncC) as molecular recognition elements (MREs) for an NMN-specific biosensor.
Methods:
- Thermal stability assays and fluorescence spectroscopy were used to analyze the binding of NMN and related metabolites to mutated PncC variants.
- The S29A and K61Q PncC variants were chosen for further analysis.
Results:
- Both S29A and K61Q PncC variants can bind to NMN, but only S29A also binds to NaMN, a product of the PncC reaction.
- K61Q PncC shows potential as a specific probe for an NMN biosensor.
Significance:
- A biosensor for NMN could provide insights into its role in cellular metabolism and disease.
- The study identified mutants of NMN deamidase that still bind NMN, making them potential candidates for an NMN biosensor.
Conclusion:
- S29A and K61Q PncC mutants have the ability to bind NMN and may be used as molecular recognition elements in an NMN-specific biosensor.
Further Considerations:
- Future studies should investigate ligand-induced conformational changes, protein stability under different conditions, and factors like pH, temperature, ionic strength, and solvent polarity for biosensor design.
- Additional data and insights related to this study can be found in the supplementary materials provided online.
Author Information:
- Alessandra Camarca, S.D., and N.R. were involved in project conceptualization, formal analysis, and funding acquisition.
- G.M., A.P., Alessandro Capo (A.C.), and A.A. contributed to the methodology.
- S.D. and N.R. supervised the project.
- A.C., A.P., and N.R. drafted the original manuscript, which was reviewed and edited by A.C., S.D., and N.R.
- All authors have approved the manuscript for publication.
Funding:
- The funding for this work was provided by Ministero Dell’Università e della Ricerca for the Prin 2017 CBNCYT project.
Ethical Considerations:
- The document states that the Institutional Review Board (IRB) statement is not applicable, suggesting that the research does not involve human subjects or falls outside the IRB's jurisdiction.
- Researchers should ensure that their work complies with ethical considerations, even when an IRB statement is not applicable.
Data Availability:
- The datasets used in the study can be accessed by contacting the corresponding author.
- Data availability is subject to the permission and discretion of the corresponding author.
- Interested individuals should reach out directly to the corresponding author to request access to the datasets.
Conflicts of Interest:
- The authors have declared that they have no conflicts of interest.
- This increases the credibility and objectivity of their work.
Publisher Note:
MDPI, the publisher, remains neutral regarding jurisdictional claims and institutional affiliations.
Title of paper: Characterization of Two NMN Deamidase Mutants as Possible Probes for an NMN Biosensor
Author(s): Camarca A, Minazzato G, Pennacchio A, Capo A, Amici A, D'Auria S, Raffaelli N.
Year published: 2021
Published in: Int J Mol Sci
Original article can be found here.
