A continuous spectrophotometric assay that distinguishes between phospholipase A1 and A2 activities.
作者： Meddy El Alaoui , Laurent Soulère , Alexandre Noiriel , Florence Popowycz , Abdallah Khatib
DOI: 10.1194/JLR.D065961
关键词:
摘要: A new spectrophotometric assay was developed to measure, continuously and specifically, phospholipase A1 (PLA1) or A2 (PLA2) activities using synthetic glycerophosphatidylcholines (PCs) containing α-eleostearic acid, either at the sn-1 position [1-α-eleostearoyl-2-octadecyl-rac-glycero-3-phosphocholine (EOPC)] sn-2 [1-octadecyl-2-α-eleostearoyl-rac-glycero-3-phosphocholine (OEPC)]. The substrates were coated onto wells of microtiter plates. nonhydrolyzable ether bond, with a non-UV-absorbing alkyl chain, introduced other sn prevent acyl chain migration during lipolysis. Upon enzyme action, acid is liberated then solubilized into micellar phase. PLA1 PLA2 activity measured by increase in absorbance 272 nm due transition from adsorbed soluble state. EOPC OEPC differentiate, excellent accuracy, between activity. Lecitase®, guinea pig pancreatic lipase-related protein 2 (known be enzyme), bee venom PLA2, porcine all used validate assay. Compared current assays for measuring and/or their inhibitors, development this sensitive enzymatic method, PC substrate analogs natural lipids based on UV spectroscopic properties significant improvement.
sci-hub.se 参考文章(50)
Ahmed Aloulou, Renaud Rahier, Yani Arhab, Alexandre Noiriel, Abdelkarim Abousalham, Phospholipases: An Overview Methods of Molecular Biology. ,vol. 1835, pp. 69- 105 ,(2012) , 10.1007/978-1-61779-600-5_4
H. van den Bosch, J.G.N. de Jong, A.J. Aarsman, Phospholipase A2 from rat liver mitochondria. Methods in Enzymology. ,vol. 197, pp. 365- 373 ,(1991) , 10.1016/0076-6879(91)97162-R
Stéphane Ransac, Youssef Gargouri, Frank Marguet, Gérard Buono, Christoph Beglinger, Pius Hildebrand, Hans Lengsfeld, Paul Hadváry, Robert Verger, Covalent inactivation of lipases. Methods in Enzymology. ,vol. 286, pp. 190- 231 ,(1997) , 10.1016/S0076-6879(97)86012-0
Stéphane Ransac, Margarita Ivanova, Ivan Panaiotov, Robert Verger, Monolayer Techniques for Studying Lipase Kinetics Lipase and Phospholipase Protocols. ,vol. 286, pp. 279- 302 ,(1999) , 10.1385/1-59259-581-2:279
P Hadváry, H Lengsfeld, H Wolfer, Inhibition of pancreatic lipase in vitro by the covalent inhibitor tetrahydrolipstatin. Biochemical Journal. ,vol. 256, pp. 357- 361 ,(1988) , 10.1042/BJ2560357
Josette Fauvel, Marie-José Bonnefis, Louis Sarda, Hugues Chap, Jean-Paul Thouvenot, Louis Douste-Blazy, Purification of two lipases with high phospholipase A1 activity from guinea-pig pancreas. Biochimica et Biophysica Acta. ,vol. 663, pp. 446- 456 ,(1981) , 10.1016/0005-2760(81)90173-9
DY Hui, MJ Cope, ED Labonté, H-T Chang, J Shao, E Goka, A Abousalham, D Charmot, J Buysse, The phospholipase A2 inhibitor methyl indoxam suppresses diet‐induced obesity and glucose intolerance in mice British Journal of Pharmacology. ,vol. 157, pp. 1263- 1269 ,(2009) , 10.1111/J.1476-5381.2009.00308.X
Xunjun Xiao, Leah E. Ross, Wednesday A. Sevilla, Yan Wang, Mark E. Lowe, Porcine pancreatic lipase related protein 2 has high triglyceride lipase activity in the absence of colipase Biochimica et Biophysica Acta. ,vol. 1831, pp. 1435- 1441 ,(2013) , 10.1016/J.BBALIP.2013.06.002
Renato Rosseto, Celize M. Tcacenco, Radha Ranganathan, Joseph Hajdu, Synthesis of Phosphatidylcholine Analogues Derived from Glyceric Acid: a New Class of Biologically Active Phospholipid Compounds. Tetrahedron Letters. ,vol. 49, pp. 3500- 3503 ,(2008) , 10.1016/J.TETLET.2008.03.084
Cécilia Eydoux, Josiane De Caro, Francine Ferrato, Paul Boullanger, Dominique Lafont, René Laugier, Frédéric Carrière, Alain De Caro, Further biochemical characterization of human pancreatic lipase-related protein 2 expressed in yeast cells. Journal of Lipid Research. ,vol. 48, pp. 1539- 1549 ,(2007) , 10.1194/JLR.M600486-JLR200