(Nicotinamide adenine dinucleotide) NAD Metabolites Analysis Service

What Is Nicotinamide Adenine Dinucleotide (NAD)

Nicotinamide adenine dinucleotide (NAD) is a coenzyme, which can be found in all living cells. NAD consists of two nucleotides joined through their phosphate groups and it is a dinucleotide, in which the one nucleotide contains an adenine base and the other nicotinamide. There are two different NAD forms exist naturally, the one is an oxidized form, which is abbreviated as NAD+ and the other is a reduced form, which is abbreviated as NADH.

What Is Nad Synthesis Pathway

In both prokaryotic and eukaryotic systems, NAD is synthesized through two pathways: de novo pathway and salvage pathway. In the de novo pathway, NAD is generated from tryptophan through quinolinic acid (QA) and nicotinic acid (NA). In salvage pathway, NAD is synthesized by recycling degraded NAD products such as Nam and nicotinamide. Both pathways play essential roles in cell growth. Under normal physiological condition, the salvage pathway plays a more important role than de novo pathway in NAD synthesis. In yeast, the de novo pathway is making of six enzymatic steps and one non-enzymatic reaction. In the last enzymatic reaction, quinolinate is converted to nicotinic acid mononucleotide by a quinolinate phosphoribosyl transferase encoded by the BNA6/ QPT1 gene. This last enzymatic reaction is the converge point of the de novo pathway and the salvage pathway.

Physiological Functions of NAD

NAD takes part in many physiological processes, such as energy metabolism regulation, DNA repair and transcription. Besides acting as a coenzyme, NAD also serves as a substrate such as substrate for NAD-dependent DNA ligases, NAD-dependent oxidoreductases and NAD-dependent deacetylases. At the same time, the reduced form of NAD, NADH, serves as a substrate for the NADH dehydrogenase in the mitochondrial respiratory chain to transfers electrons to coenzyme Q and generate NAD. Calorie restriction accomplished by glucose limitation in wild-type Saccharomyces cerevisiae can extend replicative lifespan of cells and this process relies on Sir2 and the NAD+ salvage enzymes, nicotinic acid phosphoribosyl transferase and nicotinamidase. Together with glutathione, the derivative of NAD, the NADPH coenzyme, maintains the intracellular redox state and is also involved in many assimilatory pathways. To maintain the proper redox state, NADH needs to be constantly re-oxidised. Mostly, NAD is converted to NADH mostly in catabolic reactions such as glycolysis and TCA cycle. However, both cytosolic and mitochondrial NADH are re-oxidised mainly by the respiratory chain. Since the inner mitochondrial membrane is impermeable for NAD and NADH, there are several shuttle systems to transport permeable redox equivalents across this barrier. Ethanol-acetaldehyde shuttle is one example.

Though the changes of NAD+ to nicotinamide ratio and the NAD+ to NADH ratio can be anticipated through models to related the effects of calorie restriction. However, the putative alterations of NAD+ metabolism require a sensitive and reliable qualitative and quantitative analysis of NAD+ metabolites.

What Are The Pertinent Small Molecules Within The NAD Metabolic Pathway?

The NAD metabolic pathway encompasses a spectrum of small molecules, prominently featuring the oxidized form of nicotinamide adenine dinucleotide (NAD+), its reduced counterpart nicotinamide adenine dinucleotide (NADH), the oxidized form of nicotinamide adenine dinucleotide phosphate (NADP+), the reduced form of nicotinamide adenine dinucleotide phosphate (NADPH), nicotinamide mononucleotide (NMN), nicotinamide (NAM), nicotinamide riboside (NAR), and adenosine triphosphate (ATP). Notably, nicotinamide adenine dinucleotide (NAD+), alternatively referred to as coenzyme I, plays an indispensable role as a coenzyme in redox processes, actively participating in diverse physiological processes such as DNA repair, cellular substance metabolism, and energy synthesis.

NAD Metabolites Analysis Solution

Creative Proteomics presents a cutting-edge analytical solution tailored to the study of NAD metabolism. This comprehensive platform leverages state-of-the-art GC/MS technology (Agilent 7890A/5975C) and ACQUITY UPLC/TripleQuad5500 instrumentation (Waters/AB Sciex) to facilitate both quantitative and qualitative assessments of a wide spectrum of small molecules involved in the NAD metabolic pathways. Central to this platform is the incorporation of standard reference materials and isotopic standards, enabling precise quantification of nearly 95% of the analytes under investigation.

Sample Requirement

Serum, plasma, urine, bile, and bile acids;

Animal tissues such as cells, liver, brain tissues, etc., and feces;

Plants, yeast, microorganisms, etc.

Blood samples, bile, etc.: 10 microliters

Various tissues: 10 milligrams

Feces, etc.: 10 milligrams

Delivery

A detailed technical report will be provided at the end of the whole project, including the experiment procedure, MS/MS instrument parameters

Analytes are reported as uM or ug/mg (tissue), and CV's are generally<10%

The name of the analytes, abbreviation, formula, molecular weight and CAS# would also be included in the report.

With integrated set of separation, characterization, identification and quantification systems featured with excellent robustness & reproducibility, high and ultra-sensitivity, Creative Proteomics provides reliable, rapid and cost-effective NAD metabolites targeted metabolomics services.