Browse > Article

Advanced Lipid Extraction Method for the Determination of the Phospholipase D Activity  

Yon, Chang-Suek (Institute of Biomedical Science and Department of Biochemistry, College of Medicine, Hanyang University)
Lee, Min-Ho (Department of Internal Medicine, College of Medicine, Hanyang University)
Oh, Doo-Yi (Institute of Biomedical Science and Department of Biochemistry, College of Medicine, Hanyang University)
Kim, Dal-Soo (Department of Neurosurgery, Uijongbu St. Mary′s Hospital, The Catholic University of Korea, College of Medicine, PaiChai University)
Lee, Ki-Sung (The Research Center for Bio-Medicinal Resources, PaiChai University)
Han, Joong-Soo (Institute of Biomedical Science and Department of Biochemistry, College of Medicine, Hanyang University, The Research Center for Bio-Medicinal Resources, PaiChai University)
Publication Information
Archives of Pharmacal Research / v.26, no.6, 2003 , pp. 478-481 More about this Journal
Abstract
Phospholipase D is a ubiquitous enzyme that plays an important role in various lipid mediated cellular signaling pathways and produces rare phospholipids, phosphatidylethanol or phosphatidylbutanol, instead of phosphatidic acid with unique catalytic activity transphosphatidylation in the presence of primary alcohols. The reaction products, phosphatidylethanol or phosphatidylbutanol are used as markers of in vitro phospholipase D activity in many studies. For the sensitive detection of the phospholipase D products, we developed an advanced lipid extraction method that facilitates recovery of the compounds. With the new method, the activity change of phosaholipase D by agonists could be detected more easily and the recovery rate was also increased. The increase of detected enzyme activity change was about double fold compared to the conventional lipid extraction method. This method provides selective force for the phospholipase D products in the extraction procedure.
Keywords
Phospholipase D; Phosphatidylethanol; Phosphatidylbutanol; Enzyme activity; Lipid extraction; HPLC;
Citations & Related Records

Times Cited By Web Of Science : 2  (Related Records In Web of Science)
Times Cited By SCOPUS : 2
연도 인용수 순위
1 Exton, J., Phospholipase D. Biochim. Biophys. Acta, 1436, 105-115 (1998)   DOI   PUBMED   ScienceOn
2 Gunnarsson, T., Karlsson, A., Hansson, P., Johnson, G., Alling, C., and Odham, G., Determination of phosphatidylethanol in blood from alcoholic males using high-performance liquid chromatography and evaporative light scattering or electrospray mass spectrometric detection. J. Chromatogr. B., 705(2), 243-249 (1998)   DOI   ScienceOn
3 Varga, A., Hansson, P., Johnson, G., and Alling, C., Normalization rate and cellular localization of phosphatidylethanol in whole blood from chronic alcoholics. Clin. Chim. Acta, 299(1-2), 141-150 (2000)   DOI   PUBMED   ScienceOn
4 Yon, C. and Han, J. S., Analysis of trimethylsilyl derivatization products of phosphatidylethanol by GC-MS. Exp. Mol. Med., 32, 243-245 (2000)   DOI   PUBMED   ScienceOn
5 Gomez-Cambronero, J. and Jeire, P., Phospholipase D: A Novel Major Player in Signal Transduction. Cell Signal., 10(6), 387-397 (1998)   DOI   ScienceOn
6 Kobayashi, M. and Kanfer, J. N., Phosphatidylethanol formation via transphosphatidylation by rat brain synaptosomal phospholipase D. J. Neurochem., 1597-1603 (1987)   PUBMED
7 Kim, J. H., Yoon, Y. D., Shin, I. C., and Han, J. S., Effects of ceramide, the Fas signal intermediate, on apoptosis and phospholipase D activity in mouse ovarian granulosa cells in vitro. IUBMB. Life, 48, 445-452 (1999)   DOI   PUBMED
8 Oh, T.-W., Kim, J.-H., Lee, K. S., and Han, J.-S., Major house dust mite allergen, Der pI, activates phospholipase D in human peripheral blood mononuclear cells from allergic patients : Involvement of protein kinase C. Exp. Mol. Med., 32(2), 67-71 (2000)   DOI   PUBMED   ScienceOn
9 Shin, I., Kang, J. H., and Han, J.-S., Changes of phospholipase D activity in TNF-á and anti-Fas/Apo1 monoclonal antibody induced apoptosis in HL-60 and A20 cells. Exp. Mol. Med., 30(1), 21-27 (1998)   DOI   PUBMED   ScienceOn
10 Bligh, K. M. and Dyer, W. J., A rapid method of total lipid extraction and purification. Can. J. Biochem. Physiol., 37, 911-917 (1959)   DOI   PUBMED
11 Kemken, D., Mier, K., Katus, H.A., Richardt, G., and Kurz, T., A HPLC-fluorescence detection method for determination of cardiac phospholipase D activity in vitro. Anal. Biochem., 286, 277-281 (2000)   DOI   ScienceOn
12 Park, H. S., Lee. S. Y., Kim, Y. H., Kim, J. Y., Lee, S. J., and Choi, M., Membrane perturbation by mastoparan 7 elicits a broad alteration in lipid composition of L1210 cells., Biochim. Biophys. Acta, 1484(2-3), 151-162 (2000)   DOI   PUBMED   ScienceOn