Pantothenate & CoA Biosynthetic Metabolite Analysis

What is Pantothenic Acid and Coenzyme A Biosynthesis?

Pantothenate and CoA (Coenzyme A) are essential molecules involved in various metabolic processes within cells. Pantothenate, also known as vitamin B5, serves as a precursor molecule for the biosynthesis of CoA. CoA, on the other hand, is a coenzyme that plays a critical role in energy metabolism and is involved in numerous biochemical reactions.

Pantothenate is synthesized through a series of enzymatic reactions known as the pantothenate biosynthetic pathway. It starts with the condensation of pantoate (beta-alanine) and β-alanine, followed by subsequent modifications to form pantothenate. Pantothenate is then converted into CoA through a two-step process involving the addition of an adenosine monophosphate (AMP) group and a cysteine moiety.

CoA acts as a carrier molecule, playing a central role in various metabolic pathways. It functions by binding to acyl groups derived from fatty acids, amino acids, and carbohydrates, facilitating their transport and metabolism within the cell. CoA is crucial for energy production through the citric acid cycle (also known as the Krebs cycle) and oxidative phosphorylation. It is also involved in fatty acid synthesis, amino acid metabolism, and the synthesis of various bioactive molecules.

The biosynthesis and regulation of pantothenate and CoA are tightly controlled processes in cells. Their levels are influenced by dietary intake, cellular demand, and enzymatic activities involved in their synthesis and degradation pathways. Imbalances in pantothenate and CoA metabolism can have significant impacts on cellular functions and may be associated with metabolic disorders and diseases.

Understanding the biosynthesis and metabolism of pantothenic acid and CoA is important for elucidating cellular metabolism, energy production, and various metabolic pathways. Studying the levels and regulation of these metabolites can provide valuable insights into metabolic disorders, drug targets, and therapeutic interventions.

Creative Proteomics uses a variety of analytical techniques such as high-performance liquid chromatography (HPLC), mass spectrometry (MS), and stable isotope tracking to analyze and quantify pantothenic acid and CoA biosynthetic metabolites in biological samples. These methods enable measurement of their concentrations, exploration of their interactions in metabolic networks, and investigation of their role in health and disease.

Pantothenate & CoA Metabolites Assay at Creative Proteomics

Instrumentation:

• Liquid Chromatography: The assay typically utilizes a high-performance liquid chromatography (HPLC) system or an ultra-high-performance liquid chromatography (UHPLC) system for chromatographic separation. Examples of commonly used instruments include Agilent 1200 series HPLC or Thermo Scientific Vanquish UHPLC.
• Mass Spectrometer: The liquid chromatography system is coupled with a mass spectrometer. The mass spectrometer commonly employed is a triple quadrupole mass spectrometer, such as AB Sciex Triple Quad™ 5500 or Thermo Scientific TSQ series, or a hybrid quadrupole-time-of-flight (Q-TOF) mass spectrometer, such as Waters Xevo G2-XS Q-TOF or Bruker Impact II Q-TOF.

Workflow for the pantothenate and CoA metabolites assay

• Sample preparation: Extract the metabolites from the biological samples (plasma, urine, tissue, etc.) using suitable extraction techniques.
• Chromatographic separation: Inject the extracted samples into an LC system for separation using a specific chromatographic column and optimized conditions.
• Mass spectrometry: Couple the LC system with a mass spectrometer equipped with an electrospray ionization (ESI) source to detect and quantify the metabolites.
• Calibration standards: Prepare a series of standards with known concentrations to generate a calibration curve for accurate quantification.
• Quality control samples: Include quality control samples at different concentration levels to ensure assay accuracy and precision.
• Data acquisition and analysis: Acquire mass spectrometry data, process it using specialized software for peak detection and quantification, and calculate metabolite concentrations.
• Data interpretation and reporting: Analyze the results, compare metabolite concentrations with reference ranges or baseline levels, and prepare a report summarizing the assay findings.

The overall workflow of pantothenate and CoA biosynthesis metabolites analysis

List of Partial Detectable Pantothenate And CoA Biosynthesis Related Metabolites at Creative Proteomics

Sample Requirements

Creative Proteomics can analyze a wide range of biological materials including but not limited to cells and solid tissues from animals. If you need transport your samples to us, please follow the following requirements for different types of samples:

• Blood/plasma: 500ul/sample
• Urine: 1ml/sample
• Tissue: 200mg/sample
• Cells: 1x10⁷/sample
• Feces: 500mg/sample

At least 6 biological replicates per group for cellular/microbial samples.

At least 10 biological replicates per group for model animal/plant samples.

Clinical samples generally have at least 50 biological replicates per group.

Shipment condition: dry ice

Report Delivery

• Experimental procedures
• The parameters of liquid chromatograph and mass spectrometer
• Purity analysis report
• MS raw data files and MS data quality inspection
• Absolute quantitative analysis data

Based on advanced mass spectrometry platform, professional bioinformatics software and experienced technicians and scientists, Creative Proteomics provides customers with rapid experimental process and easy-to-read reports for pantothenate and coenzyme a biosynthesis-related metabolite detection and analysis services to accelerate your scientific research.

References

1. Czumaj Aleksandra, Szrok-Jurga Sylwia, Hebanowska Areta. et al. The Pathophysiological Role of CoA. Int J Mol Sci. 2020.21:9057.
2. Leonardi Roberta, Jackowski Suzanne. Biosynthesis of Pantothenic Acid and Coenzyme A. EcoSal Plus. 2007.2:10.