Mevalonate pathway, also known as HMG-CoA reductase pathway or the isoprenoid pathway, is an important metabolic pathway essential in eukaryotes, archaea, and some bacteria. The mevalonate pathway is widely studied. This pathway is widely known being involved in cardiovascular diseases. Recently, a large number of experimental and clinical studies show that inhibition of non-sterol isoprenoids has important therapeutical implications. By synthesizing sterol isoprenoids, such as cholesterol, and non-sterol isoprenoids, this pathway plays an essential role in various cellular processes. Starting with acetyl-CoA, the mevalonate pathway ends with two five-carbon building blocks names as isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) used for making isoprenoids like cholesterol, coenzyme Q10, and all steroid hormones. This pathway, specifically HMG-CoA reductase in the mevalonate pathway, is a well-known target of a class of cholesterol lowering drugs—statins. Statins inhibits the activity of HMG-CoA reductase, and thereby inhibits the generation of cholesterol- one kind of isoprenoids.
The mevalonate pathway starts with the two consecutive enzyme catalyzed steps conversion from three acetyl-CoAs into 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA). The rate-limiting enzyme of the pathway, HMG-CoA reductase is then converts the HMG-CoA to mevalonate (MVA). 5-pyrophosphomevalonate (MVAPP) is generated after MVA is phosphorylated twice. The decarboxylation of MVAPP leads to the formation of IPP.
The mevalonate pathway plays a key role in cellular metabolism and is responsible for conversion of acetyl-CoA to isopentenyl 5-diphosphate, which is the precursor of various polyisoprenoid metabolites and natural products. These polyisoprenoid metabolites and natural products act a number of different functions. Besides generating various sterols for the synthesis of hormones, bile acids and oxysterols, the pathway also generates a wide range of non-sterol isoprenoids with various structures and functions, which are derived from isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP). Heme A acts in the mitochondrial respiratory chain, side chains of ubiquinone-10, isopentenyl-tRNA involved in protein translation, dolichol essential for protein glycosylation are examples of those non-sterol isoprenoids. These non-sterol isoprenoids are potential therapeutic targets for many diseases such as oncology, autoimmune disorders and Alzheimer disease.
In literature, a large number of detection methods for intermediates involved in the mevalonate pathway have been described. However, only one intermediate can be determined in most of these described methods, such as MVA in human urine and plasma and DMAPP in plant leaves. Though simultaneous determination of FPP and GGPP in rat liver and cultured NIH3T3 cells has been described, the simultaneous determination of all the intermediates in the mevalonate pathway remains a challenge. That’s because the intermediates involved in the mevalonate pathway are dramatically different in structure and physical properties. McCaskill and Croteau described a simultaneous radiolabeled quantification procedure for the analysis of all 11 intermediates of the mevalonate pathway in plant cells.
With no use of radioactive or fluorescent compounds, here Creative Proteomics develops a sensitive high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the direct detection and quantification of all intermediates in the mevalonate pathway.
Platform
- UPLC-MRM/MS
Summary
- Identification and quantification of intermediates in the mevalonate pathway by mixed organic solvent extraction in cells or tissue.
Sample Requirement
- Normal Volume: 200ul serum/plasma; 200 mg tissue, 2e7 cells
- Minimal Volume: 50ul serum/plasma; 50 mg tissue, 5e6 cells
Report
- 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.
| Mevalonate Pathway Metabolites Quantified in This Service | ||
|---|---|---|
| Acetoacetyl-CoA | Acetyl-CoA | Dimethylallyl-P-P |
| Geranylgeranyl-P-P | Geranyl-P-P | HMG-CoA |
| Isopentenyl-P-P | Mevalonate | Thamine pyrophosphate |
| Thiamine monophosphate | (E,E)-Farnesyl-P-P | 5-P-mevalonate |
| 5-P-P-mevalonate | ||
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 mevalonate pathway targeted metabolomics services.