caa a22 4500 465766463 CHVBK 20180323111915.0 cr unu---uuuuu 170327e19900801xx s 000 0 eng 10.1007/BF02538084 doi (NATIONALLICENCE)springer-10.1007/BF02538084 Hydrolysis of fluorescent pyrene-acyl esters by human pancreatic carboxylic ester hydrolase and bile salt-stimulated lipase [Elektronische Daten] [Anne Negre-Salvayre, Nehza Abouakil, Dominique Lombardo, Robert Salvayre] Fluorescent esters containing pyrenedecanoic acid (P10) or pyrenebutanoic (P4) acid (P4cholesterol, P10cholesterol, P4- and P10-containing triacylglycerols) were synthesized and used as substrates for human pancreatic carboxylic ester hydrolase and bile salt-stimulated lipase from human milk. Both enzymes were purified by immunoaffinity chromatography. All fluorescent pyrene derivatives were hydrolyzed by pancreatic carboxylic ester hydrolase and bile salt-stimulated lipase, but at different rates. The hydrolytic rates of the "short” acyl esters (P4-containing esters) were higher than those of the "long” ones (P10-containing esters). Conditions were optimized for sensitivity of the assay using fluorescent cholesteryl esters. The pH optimum was 7.5-8.0. Sodium cholate exhibited a stronger activating effect than taurocholate or taurodeoxycholate (maximal activation was achieved with 5 mmol/L cholate and with a molar ratio cholesteryl ester/cholate around 1∶10). Both pancreatic carboxylic ester hydrolase and bile salt-stimulated lipase from milk were strongly inhibited by the other amphiphiles tested, namely phosphatidylcholine and Triton X-100, and were inactivated by low concentrations (10 μmol/L) of the serine-reactive diethyl-paranitrophenyl phosphate (E600). Both enzymes were strongly inhibited by relatively low concentrations of plasma low density lipoproteins. These studies indicate that the fluorescent esters containing pyrene fatty acids can be used as substrates for assaying and investigating the properties of pancreatic carboxylic ester hydrolase as well as bile salt-stimulated lipase from milk. American Oil Chemists' Society (AOCS), 1990 Negre-Salvayre Anne Laboratoire de Biochimie, CHU Rangueil, Av. J. Poulhes, 31054, Toulouse Cedex aut Abouakil Nehza CBBM-CNRS, 13402, Marseille Cedex, France aut Lombardo Dominique CBBM-CNRS, 13402, Marseille Cedex, France aut Salvayre Robert Laboratoire de Biochimie, CHU Rangueil, Av. J. Poulhes, 31054, Toulouse Cedex aut Lipids Springer-Verlag 25/8(1990-08-01), 428-434 0024-4201 25:8<428 1990 25 11745 https://doi.org/10.1007/BF02538084 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF02538084 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Negre-Salvayre Anne Laboratoire de Biochimie, CHU Rangueil, Av. J. Poulhes, 31054, Toulouse Cedex aut NATIONALLICENCE 700 1- Abouakil Nehza CBBM-CNRS, 13402, Marseille Cedex, France aut NATIONALLICENCE 700 1- Lombardo Dominique CBBM-CNRS, 13402, Marseille Cedex, France aut NATIONALLICENCE 700 1- Salvayre Robert Laboratoire de Biochimie, CHU Rangueil, Av. J. Poulhes, 31054, Toulouse Cedex aut NATIONALLICENCE 773 0- Lipids Springer-Verlag 25/8(1990-08-01), 428-434 0024-4201 25:8<428 1990 25 11745 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer