A novel assay for measuring recombinant human lysophosphatidylcholine acyltransferase 3 activity
| العنوان: | A novel assay for measuring recombinant human lysophosphatidylcholine acyltransferase 3 activity |
|---|---|
| المؤلفون: | Xinxin Xiang, Jia-chun Hu, Qi Liu, Xinming Du, Jibin Dong, Xiang Gu, Yu Cao, Qing Zhang, Tingbo Ding, Yingxia Li, Shu-hua He, Bin Lou |
| المصدر: | FEBS Open Bio FEBS Open Bio, Vol 9, Iss 10, Pp 1734-1743 (2019) |
| سنة النشر: | 2019 |
| مصطلحات موضوعية: | 0301 basic medicine, LPCAT3, Cell, Phospholipid, General Biochemistry, Genetics and Molecular Biology, law.invention, 03 medical and health sciences, chemistry.chemical_compound, Mice, 0302 clinical medicine, law, enzyme kinetics, medicine, Animals, Humans, Enzyme kinetics, Enzyme Inhibitors, IC50, lcsh:QH301-705.5, Research Articles, sequential mechanism, Enzyme Assays, chemistry.chemical_classification, Temperature, 1-Acylglycerophosphocholine O-Acyltransferase, inhibitor screening, Hydrogen-Ion Concentration, medicine.disease, lysophosphatidylcholine acyltransferase 3, Recombinant Proteins, Mice, Inbred C57BL, Kinetics, 030104 developmental biology, medicine.anatomical_structure, Lysophosphatidylcholine, Enzyme, chemistry, Biochemistry, lcsh:Biology (General), 030220 oncology & carcinogenesis, Recombinant DNA, Steatohepatitis, Research Article |
| الوصف: | Lysophosphatidylcholine acyltransferase 3 (LPCAT3) is an important enzyme in phospholipid remodeling, a process that influences the biophysical properties of cell membranes and thus cell function. Multiple lines of evidence suggest that LPCAT3 is involved in several diseases, including atherosclerosis, non-alcoholic steatohepatitis, and carcinoma. Thus, LPCAT3 may have potential as a therapeutic target for these diseases. In the present study, we devised an assay based on reversed-phase HPLC to measure LPCAT3 activity, which may facilitate the identification of LPCAT3 inhibitors and activators. We found that optimal pH and temperature of recombinant human LPCAT3 are 6.0 and 30 °C, respectively. The enzyme Km values for substrates NBD-labelled lysophosphatidylcholine and arachidonoyl CoA were 266.84 ± 3.65 and 11.03 ± 0.51 μmol·L-1 , respectively, and the Vmax was 39.76 ± 1.86 pmol·min-1 ·U-1 . Moreover, we used our new method to determine the IC50 of a known LPCAT inhibitor, TSI-10. In conclusion, this novel assay can be used to measure the effects of compounds on LPCAT3 activity. |
| تدمد: | 2211-5463 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::9658d7736b85b634a3160ea498544a38 https://pubmed.ncbi.nlm.nih.gov/31376210 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....9658d7736b85b634a3160ea498544a38 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 22115463 |
|---|