Nucleotide Metabolic Flux Analysis

Nucleotide Metabolic Flux Analysis

Nucleotides consist of a nitrogenous base as a core, plus a five-carbon sugar and a phosphate group. There are five possible nitrogenous bases, namely adenine, guanine, cytosine, thymine and uracil. As precursors for the synthesis of nucleic acids, nucleotides have important biological functions and are involved in almost all biochemical processes in living organisms. Metabolic flux analysis (MFA) is the dynamic process of tracing metabolites over time throughout a network of metabolic reactions, using stable isotope tracing techniques. Creative Proteomics has established a MFA platform primarily based on mass spectrometry (MS) detection. We offer leading international services in MFA of multiple pathways, including nucleotide MFA.

Physiological functions of nucleotides

The major physiological functions of nucleotides in the body are:

• As raw materials for the synthesis of nucleic acids, such as the synthesis of RNA and DNA
• As a form of energy storage and supply, in addition to ATP, GTP, UTP, CTP, etc.
• Involved in the regulation of metabolic or physiological activities, such as cyclic nucleotides CAMP and cGMP as the second messenger of hormones
• Involved in the formation of enzyme coenzymes or cofactors, such as in NAD+, NADP+, FAD, FMN, CoA all contain nucleotide components
• As carriers of metabolic intermediates, such as UDP carries sugar groups, CDP carries choline, cholamine or diglycerides, adenosine carries methionine (SAM), etc.

Nucleotide metabolic flux analysis service at Creative Proteomics

Creative Proteomics is proud to offer nucleotide MFA service, which can help researchers and professionals better understand the metabolite level changes, flux distribution and turnover rates of intracellular nucleotide metabolic networks, investigate major metabolic abnormal pathways and their biological functions, and reveal their upstream and downstream inter-regulatory mechanisms. Specifically, our service includes experimental design, labeling experiments, isotopic labeling, nucleotide metabolic flux estimation, and metabolic flux data analysis. We are committed to providing a strong scientific basis for the understanding of disease mechanisms, drug target discovery and other fields.

Analytical list of nucleotides and related substances

Based on our advanced platform and experienced experts, we are able to perform quantitative and qualitative analysis of the following nucleotides and related substances, but not limited to.

Accepted samples

Cells, tissues, microbial organisms, culture media and fermentation broth

Instrument platform

Orbitrap LC-MS, UHPLC-QQQ-MS, and GC-Q-MS

Report delivery

• Experimental steps
• Relevant MS parameters
• Quantitative or quantitative information of the identified substances
• Mass spectral images
• Raw data
• Metabolic flux result and interpretation, including variance analysis, cluster analysis, correlation analysis, ROC analysis, network analysis, and other custom analysis

Creative Proteomics is a leading service provider in MFA. We can offer various ¹³C or ¹⁵N stable isotope labeled metabolic precursors, such as D-Glucose (U-¹³C₆), D-Glucose (1,2-¹³C₂), D-Glucose (1-¹³C), L-Glutamine (¹³C₅), L-Glutamine (amine-¹⁵N), L-Glutamine (1-¹³C), and L-Glutamine (5-¹³C). A range of other uniformly or selectively ¹³C/¹⁵N/D -labeled metabolites and amino-acids are also available. We will develop a detailed service agreement according to your program and needs. For more experimental technology services, please browse the rest of our website or contact us.

Reference

1. Lane, Andrew N., and Teresa WM Fan. "Regulation of mammalian nucleotide metabolism and biosynthesis." Nucleic acids research 43.4 (2015): 2466-2485.