A membrane-bound monoacylglycerol lipase (MAGL) activity, previously demonstrated in intact human erythrocytes [Boyer, Somma, Vérine, L'Hôte, Finidori, Merger and Arnaud (1981) J. Clin. Endocrinol. Metab. 53, 143-148], has now been purified to apparent homogeneity by a five-step procedure involving solubilization in CHAPS and sequential chromatographies on Sephacryl S-400, DEAE-Trisacryl, Zn(2+)-chelating Sepharose and Superose 12 columns. The purified protein has a molecular mass of 68 +/- 2 kDa, as determined by SDS/PAGE and gel filtration, suggesting that the enzyme behaves as a monomer. The concentration-dependence of MAGL activity with monooleoylglycerol, the preferred substrate showed kinetics typical of an interfacial lipolytic enzyme displaying optimal activity on emulsified substrate particles; apparent Km values were 0.27 mM and 0.49 mM for the sn-1(3)- and sn-2-isomers respectively. MAGL had no, or negligible, activity towards tri-oleoylglycerol, di-oleoylglycerol, oleoylcholesterol, oleoyl-CoA and phosphatidylcholine; it was inhibited by di-isopropylfluorophosphate, PMSF and diethyl p-nitrophenyl phosphate, suggesting that MAGL is a serine hydrolase. MAGL activity was not modified by bile salt or apolipoprotein C-II, whereas a dose-dependent inhibition was observed with apolipoprotein A-I.
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December 1995
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Research Article|
December 01 1995
Purification and properties of a monoacylglycerol lipase in human erythrocytes Available to Purchase
C Somma-Delpéro;
C Somma-Delpéro
1INSERM U. 260, Faculté de Médecine, 13385 Marseille Cédex 5, France
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A Valette;
A Valette
1INSERM U. 260, Faculté de Médecine, 13385 Marseille Cédex 5, France
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J Lepetit-Thévenin;
J Lepetit-Thévenin
1INSERM U. 260, Faculté de Médecine, 13385 Marseille Cédex 5, France
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O Nobili;
O Nobili
1INSERM U. 260, Faculté de Médecine, 13385 Marseille Cédex 5, France
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J Boyer;
J Boyer
1INSERM U. 260, Faculté de Médecine, 13385 Marseille Cédex 5, France
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A Vérine
A Vérine
1INSERM U. 260, Faculté de Médecine, 13385 Marseille Cédex 5, France
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Publisher: Portland Press Ltd
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© 1995 The Biochemical Society, London
1995
Biochem J (1995) 312 (2): 519–525.
Citation
C Somma-Delpéro, A Valette, J Lepetit-Thévenin, O Nobili, J Boyer, A Vérine; Purification and properties of a monoacylglycerol lipase in human erythrocytes. Biochem J 1 December 1995; 312 (2): 519–525. doi: https://doi.org/10.1042/bj3120519
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