Cloning, expression and characterization of mouse (Mus musculus) nicotinamide 5'-mononucleotide adenylyltransferase-2

David W. Raches, Suhong Xiao, Praveen Kusumanchi, Joel Yalowitz, Paresh Sanghani, Eric C. Long, Asok Antony, Hiremagalur N. Jayaram

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Nicotinamide adenine dinucleotide (NAD+) is synthesized by the action of nicotinamide mononucleotide adenylyltransferase (NMNAT) from NMN and ATP. The mouse homolog of NMNAT-2 (mmNMNAT-2) was cloned, expressed, and subsequently identified using MALDI-TOF in conjunction with the ProFound database. Circular dichroism analyses of recombinant mmNMNAT-2 showed α helical and β sheet secondary structures, consistent with the known structure of the human isoform. Competition experiments using mouse pancreatic tissue lysates with recombinant mmNMNAT-2 demonstrated that the activity of the expressed protein was similar to the human isoform. Immunohistochemistry of mouse embryonic tissues with hNMNAT-2 also showed a tissue- and cellular-specific expression of this isoform. Therefore, our studies demonstrate for the first time the clear biological evidence for the existence of a mouse isoform of hNMNAT-2. These studies may help in future investigations aimed at understanding the regulation of this gene and its pathway, and in turn, will spur the development of novel therapies for diseases such as cancer and diabetes since mice are the most frequently used experimental system for in vivo studies.

Original languageEnglish
Pages (from-to)718-726
Number of pages9
JournalMedicinal Chemistry
Volume7
Issue number6
StatePublished - Nov 2011

Fingerprint

Nicotinamide-Nucleotide Adenylyltransferase
Organism Cloning
Protein Isoforms
NAD
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Circular Dichroism
Adenosine Triphosphate
Immunohistochemistry
Databases

Keywords

  • Circular dichroism
  • MALDI-TOF
  • Mouse NMNAT-2
  • NAD
  • Tiazofurin
  • Tissue expression

ASJC Scopus subject areas

  • Drug Discovery

Cite this

Raches, D. W., Xiao, S., Kusumanchi, P., Yalowitz, J., Sanghani, P., Long, E. C., ... Jayaram, H. N. (2011). Cloning, expression and characterization of mouse (Mus musculus) nicotinamide 5'-mononucleotide adenylyltransferase-2. Medicinal Chemistry, 7(6), 718-726.

Cloning, expression and characterization of mouse (Mus musculus) nicotinamide 5'-mononucleotide adenylyltransferase-2. / Raches, David W.; Xiao, Suhong; Kusumanchi, Praveen; Yalowitz, Joel; Sanghani, Paresh; Long, Eric C.; Antony, Asok; Jayaram, Hiremagalur N.

In: Medicinal Chemistry, Vol. 7, No. 6, 11.2011, p. 718-726.

Research output: Contribution to journalArticle

Raches, DW, Xiao, S, Kusumanchi, P, Yalowitz, J, Sanghani, P, Long, EC, Antony, A & Jayaram, HN 2011, 'Cloning, expression and characterization of mouse (Mus musculus) nicotinamide 5'-mononucleotide adenylyltransferase-2', Medicinal Chemistry, vol. 7, no. 6, pp. 718-726.
Raches DW, Xiao S, Kusumanchi P, Yalowitz J, Sanghani P, Long EC et al. Cloning, expression and characterization of mouse (Mus musculus) nicotinamide 5'-mononucleotide adenylyltransferase-2. Medicinal Chemistry. 2011 Nov;7(6):718-726.
Raches, David W. ; Xiao, Suhong ; Kusumanchi, Praveen ; Yalowitz, Joel ; Sanghani, Paresh ; Long, Eric C. ; Antony, Asok ; Jayaram, Hiremagalur N. / Cloning, expression and characterization of mouse (Mus musculus) nicotinamide 5'-mononucleotide adenylyltransferase-2. In: Medicinal Chemistry. 2011 ; Vol. 7, No. 6. pp. 718-726.
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