Advanced lipid extraction method for the determination of the phospholipase D activity.
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 phospholipase 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.
Yon
Changsuek
C
Institute of Biomedical Science and Department of Biochemistry, College of Medicine, Hanyang University, 17 Seoul 133-792, Korea.
Lee
Min Ho
MH
Oh
Doo-Yi
DY
Kim
Dal-Soo
DS
Lee
Ki Sung
KS
Han
Joong-Soo
JS
D045744:Cell Line, Tumor / Q000201:enzymology; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004789:Enzyme Activation / Q000502:physiology; D006801:Humans; D008055:Lipids / Q000032:analysis*; D010739:Phospholipase D / Q000032:analysis* / Q000378:metabolism*
D008055:Lipids; D010739:Phospholipase D