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Quantifying the cellular NAD+ metabolome using a tandem liquid chromatography mass spectrometry approach.
Bustamante, Sonia; Jayasena, Tharusha; Richani, Dulama; Gilchrist, Robert Bruce; Wu, Lindsay E; Sinclair, David A; Sachdev, Perminder Singh; Braidy, Nady.
Afiliación
  • Bustamante S; Mark Wainwright Analytical Centre, University of New South Wales, Sydney, Australia.
  • Jayasena T; Faculty of Medicine, School of Psychiatry, Centre for Healthy Brain Ageing, University of New South Wales Sydney, Sydney, Australia.
  • Richani D; Faculty of Medicine, School of Women's and Children's Health, University of New South Wales Sydney, Sydney, Australia.
  • Gilchrist RB; Faculty of Medicine, School of Women's and Children's Health, University of New South Wales Sydney, Sydney, Australia.
  • Wu LE; Department of Pharmacology, School of Medical Sciences, University of New South Wales Sydney, Sydney, NSW, 2052, Australia.
  • Sinclair DA; Department of Pharmacology, School of Medical Sciences, University of New South Wales Sydney, Sydney, NSW, 2052, Australia.
  • Sachdev PS; Department of Genetics, Paul F. Glenn Center for the Biology of Aging, Harvard Medical School, Boston, MA, 02115, USA.
  • Braidy N; Faculty of Medicine, School of Psychiatry, Centre for Healthy Brain Ageing, University of New South Wales Sydney, Sydney, Australia.
Metabolomics ; 14(1): 15, 2017 12 23.
Article en En | MEDLINE | ID: mdl-30830318
ABSTRACT

INTRODUCTION:

Nicotinamide adenine dinucleotide (NAD+) is an essential pyridine nucleotide that serves as a key hydride transfer coenzyme for several oxidoreductases. It is also the substrate for intracellular secondary messenger signalling by CD38 glycohydrolases, DNA repair by poly(adenosine diphosphate ribose) polymerase, and epigenetic regulation of gene expression by a class of histone deacetylase enzymes known as sirtuins. The measurement of NAD+ and its related metabolites (hereafter, the NAD+ metabolome) represents an important indicator of cellular function.

OBJECTIVES:

A study was performed to develop a sensitive, selective, robust, reproducible, and rapid method for the concurrent quantitative determination of intracellular levels of the NAD+ metabolome in glial and oocyte cell extracts using liquid chromatography coupled to mass spectrometry (LC/MS/MS).

METHODS:

The metabolites were separated on a versatile amino column using a dual HILIC-RP gradient with heated electrospray (HESI) tandem mass spectrometry detection in mixed polarity multiple reaction monitoring mode.

RESULTS:

Quantification of 17 metabolites in the NAD+ metabolome in U251 human astroglioma cells could be achieved. Changes in NAD+ metabolism in U251 cell line, and murine oocytes under different culture conditions were also investigated.

CONCLUSION:

This method can be used as a sensitive profiling tool, tailoring chromatography for metabolites that express significant pathophysiological changes in several disease conditions and is indispensable for targeted analysis.
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Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Asunto principal: Extractos Celulares / NAD Límite: Animals / Humans Idioma: En Revista: Metabolomics Año: 2017 Tipo del documento: Article País de afiliación: Australia
Texto completo
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Asunto principal: Extractos Celulares / NAD Límite: Animals / Humans Idioma: En Revista: Metabolomics Año: 2017 Tipo del documento: Article País de afiliación: Australia