Biochemical and Biophysical Research Communications
Hydrolysis of a fluorescent phospholipid substrate by phospholipase A2 and lipoprotein lipase
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Cited by (59)
HPLC-fluorimetric assay of phospholipase A <inf>2</inf>. Application to biological samples with high protein content and various reaction conditions
2011, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life SciencesCitation Excerpt :The combination of these methods with lipid extraction and separation of the derivatized reaction products by HPLC [26] are often necessary to avoid limitations related to intrinsic properties of the biological samples [27]. Fluorescent NBD-PC derivatives have been utilized for the determination of authentic PLA2 and lipoprotein lipase preparations [28]. Based on this work we have previously reported a real-time fluorimetric assay for total PLA2 activity, with preference to long acyl chains, and PAF-AH, with preference to acetyl-groups and short, oxidatively fragmented acyl chains at the sn-2 position.
Methods for detection and characterization of lipases: A comprehensive review
2009, Biotechnology AdvancesCitation Excerpt :These molecules are chemically unrelated to TAGs and can be hydrolyzed by non-specific esterases. A second group of fluorogenic substrates consists of dansyl-phosphatidylethanolamine (Johnson et al., 1980) and NBD-phospholipids (Wittenauer et al., 1984a,b) which were reported as substrate for the phospholipase activity of lipoprotein lipase. The pyrene group was also used as a fluorophore in continuous assays of lecithin: cholesterol acyltransferase (Bonelli and Jonas, 1992), cholesteryl ester hydrolase (Joutti et al., 1985), phospholipase C (Thuren and Kinnunen, 1991), and phospholipases A (Hendrickson and Rauk, 1981; Thuren et al., 1988).
Physalin B inhibits Rhodnius prolixus hemocyte phagocytosis and microaggregation by the activation of endogenous PAF-acetyl hydrolase activities
2009, Journal of Insect PhysiologyCitation Excerpt :Arachidonic acid (5,8,11,14-eicosatetranoic acid) purchased from Sigma was dried, suspended in sodium bicarbonate 150 mM, and diluted in Grace's Insect Medium (GIM, Gibco-BRL) plus 0.25% (w/v) BSA dissolved in insect saline (1:1) to give a final concentration of 10 μM. The phospholipid substrates 1-palmitoyl-2-[12-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]dodecanoyl]-sn-glycero-3-phosphocholine (C12-NBD-PC, Avanti Polar Lipids) and 1-palmitoyl-2-{6-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]hexanoyl}-sn-glycero-3-phosphocoline (C6-NBD-PC, Avanti Polar Lipids) were prepared above their critical micellar concentrations (Wittenauer et al., 1984). Vesicles were formed by the addition of ethanol (75 μl) to 2 mg of substrate followed by flushing with a Hamilton syringe into 2 mL of 0.16 M KCl (Batzri and Korn, 1973).
Sensitive assay for hormone-sensitive lipase using NBD-labeled monoacylglycerol to detect low activities in rat adipocytes
2005, Journal of Lipid ResearchCharacterisation of a plasma membrane-associated phospholipase A<inf>2</inf> activity increased in response to cold acclimation
2002, Journal of Plant Physiology
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Present address: The Second Department of Internal Medicine, Chiba University School of Medicine, Chiba, Japan