Acyl-coenzyme A (Acyl-CoA) is a family of coenzymes involved in the metabolism of fatty acids. It is a temporary compound formed when CoA attaches to the end of a long-chain fatty acid in living cells. This compound undergoes beta oxidation, forming at least one molecules of acetyl-CoA. This, in turn, enters the citric acid cycle, finally forming several molecules of ATP.
The oxidation of fatty acids in the mitochondria gives rise to the generation of energy to be utilized when there is an increased requirement for energy such as illness, fasting, and muscular exertion. The biological process of fatty acid oxidation (FAO) includes the carnitine cycle, the betaoxidation cycle, the electron-transport chain, and, in liver, ketone synthesis. Free fatty acids are activated to their CoA esters in the cytosol at the outer mitochondrial membrane. Long-chain (C16 and C18) fatty acyl-CoAs enter the mitochondria as acylcarnitines and are reconverted back to their respective acyl-CoA's at the mitochondrial membrane.
With each step of FAO, fatty acyl-CoAs are chain-shortened by two carbons until fully converted to acetyl-CoA. Transfer of energy released during beta-oxidation to the electron transport chain results in the generation of adenosine triphosphate (ATP). In the liver, most of the acetyl-CoA is applied to synthesize ketone bodies (3-hydroxybutyrate and acetoacetate), which can be utilized as fuel by tissues that are unable to oxidize fatty acids, most remarkably, the brain. Fatty acids can be directly used by tissues such as cardiac and skeletal muscle as a source of energy. Disorders in FAO can result in various phenotypes including brain damage, hypoglycemia, kidney dysfunction, liver disease, and damage to cardiac and skeletal muscle.
Early detection of FAO disorders is important to ensure that the proper evaluation and management are carried out in a timely fashion. The major accumulating metabolites in fatty acid oxidation defects are intramitochondrial acyl-CoAs. Usually, secondary metabolites such as acylcarnitines, acylglycines and dicarboxylic acids are measured to study these defects. Most published approaches involve HPLC separations and all previously published approaches do not detect a complete range of acyl-CoA species, primarily due to hydrophobicity differences between long-, medium- and short-chain acyl-CoA species. An analytical approach has been developed by Andrew A. Palladino to directly measure diverse fatty acyl-CoA species of all chain lengths using flow-injection tandem MS. This method was evaluated using liver from the short-chain-acyl-CoA dehydrogenase deficient mouse and tested with the medium- and short-chain-3-hydroxyacyl-CoA dehydrogenase knock out mouse.
Currently, a reliable and reproducible method using highly sensitive LC-MS/MS platform for the identification and quantification of diverse acyl-CoA species in different sample types has been established by the scientists at Creative Proteomics, which can satisfy the needs of academic and industrial study in your lab.
Platform
- LC-MS/MS
Summary
Identification and quantification of diverse acyl-CoA species by mixed organic solvent extraction. Then, extracts are dried, re-suspended for LC/MS separation and measured on a LC-MS mass spectrometer using MRM methods.
Sample Requirement
- Normal Volume: 200 uL plasma, 20 mg tissue, 1e7 cells
- Minimal Volume: 50 uL, 5 mg tissue, 6e6 cells
Report
- A full report including all raw data, MS/MS instrument parameters and step-by-step calculations will be provided (Excel and PDF formats).
- Analytes are reported as uM, with CV's generally ~10%.
| Representative acyl-CoA species that can be measured by targeted LC-MS/MS | ||
|---|---|---|
| Arachidyl-CoA | Arachidonyl-CoA | Behenyl-CoA |
| Cerotyl-CoA | Linoleoyl-CoA | Lignoceryl-CoA (Tetracosahexaenoyl) |
| Oleoyl-CoA | Palmitoyl-CoA | Palmitoleoyl-CoA |
| Stearoyl-CoA | ||
Ordering Procedure:
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Staffed by experienced biological scientists, Creative Proteomics can provide a wide range of services ranging from the sample preparation to the lipid extraction, characterization, identification and quantification. We promise accurate and reliable analysis, in shorter duration of time! You are welcome to discuss your project with us.