Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1995 Dec 1;312(Pt 2):519–525. doi: 10.1042/bj3120519

Purification and properties of a monoacylglycerol lipase in human erythrocytes.

C Somma-Delpéro 1, A Valette 1, J Lepetit-Thévenin 1, O Nobili 1, J Boyer 1, A Vérine 1
PMCID: PMC1136293  PMID: 8526865

Abstract

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.

Full text

PDF
520

Images in this article

522Figure 3
on p.522

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arnaud J., Boyer J. Hydrolysis and uptake of an aliphatic fatty ester by whole isolated fat cells. Biochim Biophys Acta. 1977 Mar 25;486(3):462–469. doi: 10.1016/0005-2760(77)90096-0. [DOI] [PubMed] [Google Scholar]
  2. Arnaud J., Boyer J. Identification of an acylcholesterol lipase activity in human adipose tissue. Biochim Biophys Acta. 1974 Jan 23;337(1):165–168. doi: 10.1016/0005-2760(74)90052-6. [DOI] [PubMed] [Google Scholar]
  3. Arnaud J., Boyer J. Lipolytic activity of whole isolated liver cells in aqueous suspension. Biochim Biophys Acta. 1976 Mar 26;424(3):460–468. doi: 10.1016/0005-2760(76)90035-7. [DOI] [PubMed] [Google Scholar]
  4. Arnaud J., Nobili O., Boyer J. Characterization of a monoester lipase active as membrane-bound enzyme in rat erythrocytes. FEBS Lett. 1979 Mar 1;99(1):43–46. doi: 10.1016/0014-5793(79)80244-6. [DOI] [PubMed] [Google Scholar]
  5. Arnaud J., Nobili O., Boyer J. Characterization of a monoester lipase active as membrane-bound enzyme in rat erythrocytes. FEBS Lett. 1979 Mar 1;99(1):43–46. doi: 10.1016/0014-5793(79)80244-6. [DOI] [PubMed] [Google Scholar]
  6. Arvidsson E. O., Belfrage P. Monoglyceride-protein interaction. The binding of monoolein to native human serum albumin. Acta Chem Scand. 1969;23(1):232–236. doi: 10.3891/acta.chem.scand.23-0232. [DOI] [PubMed] [Google Scholar]
  7. Benkirane M., Meignen J. M., Mercier L., Boyer J., Verine A. Lipoprotein lipase in rat heart--III. Effects of sex steroid hormones on tri-, di- and monoacylglycerol lipase activities in post-heparin effluents. Comp Biochem Physiol B. 1989;94(1):27–30. doi: 10.1016/0305-0491(89)90005-9. [DOI] [PubMed] [Google Scholar]
  8. Berglund L., Khoo J. C., Jensen D., Steinberg D. Resolution of hormone-sensitive triglyceride/diglyceride lipase from monoglyceride lipase of chicken adipose tissue. J Biol Chem. 1980 Jun 10;255(11):5420–5428. [PubMed] [Google Scholar]
  9. Biale Y., Gorin E., Shafrir E. Characterization of tissue lipolytic and esterolytic activities cleaving full and partial glycerides. Biochim Biophys Acta. 1968 Jan 10;152(1):28–32. doi: 10.1016/0005-2760(68)90005-2. [DOI] [PubMed] [Google Scholar]
  10. Boyer J., Somma C., Vérine A., L'Hôte C., Finidori J., Merger C., Arnaud J. Human erythrocyte monoester lipase: characterization and radiochemical assay of the cell-bound enzyme in normal subjects. J Clin Endocrinol Metab. 1981 Jul;53(1):143–148. doi: 10.1210/jcem-53-1-143. [DOI] [PubMed] [Google Scholar]
  11. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  12. Bry K., Andersson L. C., Kuusi T., Kinnunen P. K. Monoacylglycerol hydrolase in human platelets. Biochim Biophys Acta. 1979 Oct 26;575(1):121–127. doi: 10.1016/0005-2760(79)90137-1. [DOI] [PubMed] [Google Scholar]
  13. CARROLL K. K. Separation of lipid classes by chromatography on Florisil. J Lipid Res. 1961 Apr;2:135–141. [PubMed] [Google Scholar]
  14. Cohen J., Somma-Delpero C., Verine A., Codaccioni J. L., Boyer J. Increased monoester lipase activity in red blood cells during hyperthyroidism. J Endocrinol. 1986 Mar;108(3):357–359. doi: 10.1677/joe.0.1080357. [DOI] [PubMed] [Google Scholar]
  15. De Jong B. J., Hülsmann W. C. Monoacylglycerol hydrolase activity of isolated rat small intestinal epithelial cells. Biochim Biophys Acta. 1978 Jan 27;528(1):36–46. doi: 10.1016/0005-2760(78)90050-4. [DOI] [PubMed] [Google Scholar]
  16. Delpéro C., Gastaldi M., Vérine A., Campistron M., Boyer J. Increased monoester lipase activity of red blood cells in alcoholism. Alcohol Clin Exp Res. 1986 Dec;10(6):602–605. doi: 10.1111/j.1530-0277.1986.tb05152.x. [DOI] [PubMed] [Google Scholar]
  17. Eisenberg S. High density lipoprotein metabolism. J Lipid Res. 1984 Oct;25(10):1017–1058. [PubMed] [Google Scholar]
  18. Gastaut J. A., Vérine A., Crcassonne Y., Boyer J. Monoester lipase activity in the red cells of patients with various blood disorders. Br J Haematol. 1982 Sep;52(1):127–130. doi: 10.1111/j.1365-2141.1982.tb03869.x. [DOI] [PubMed] [Google Scholar]
  19. Giudicelli H., Boyer J. Effects of glycerol on human adipose tissue triglyceride lipase activity. J Lipid Res. 1973 Sep;14(5):592–595. [PubMed] [Google Scholar]
  20. Hee-Cheong M., Severson D. L. Properties of monoacylglycerol lipase in rabbit aorta. Lipids. 1987 Dec;22(12):999–1004. doi: 10.1007/BF02536439. [DOI] [PubMed] [Google Scholar]
  21. Jensen G. L., Daggy B., Bensadoun A. Triacylglycerol lipase, monoacylglycerol lipase and phospholipase activities of highly purified rat hepatic lipase. Biochim Biophys Acta. 1982 Mar 12;710(3):464–470. doi: 10.1016/0005-2760(82)90130-8. [DOI] [PubMed] [Google Scholar]
  22. Kuusi T., Bry K., Nikkilä E. A., Kinnunen P. K. Modification of the substrate specificity of rat hepatic lipase by collagenase treatment. Med Biol. 1979 Jun;57(3):192–195. [PubMed] [Google Scholar]
  23. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  24. Lehner R., Kuksis A. Purification of an acyl-CoA hydrolase from rat intestinal microsomes. A candidate acyl-enzyme intermediate in glycerolipid acylation. J Biol Chem. 1993 Nov 25;268(33):24726–24733. [PubMed] [Google Scholar]
  25. Liang H. Q., Rye K. A., Barter P. J. Dissociation of lipid-free apolipoprotein A-I from high density lipoproteins. J Lipid Res. 1994 Jul;35(7):1187–1199. [PubMed] [Google Scholar]
  26. Lombardo D., Fauvel J., Guy O. Studies on the substrate specificity of a carboxyl ester hydrolase from human pancreatic juice. I. Action on carboxyl esters, glycerides and phospholipids. Biochim Biophys Acta. 1980 Jan 11;611(1):136–146. doi: 10.1016/0005-2744(80)90049-2. [DOI] [PubMed] [Google Scholar]
  27. MORRISON W. R., SMITH L. M. PREPARATION OF FATTY ACID METHYL ESTERS AND DIMETHYLACETALS FROM LIPIDS WITH BORON FLUORIDE--METHANOL. J Lipid Res. 1964 Oct;5:600–608. [PubMed] [Google Scholar]
  28. Marteau C., Quibel J. R., Le Petit-Thèvenin J., Boyer J., Gérolami A. Lipolytic activities of freshly isolated rat liver parenchymal cells. Life Sci. 1988;42(5):533–538. doi: 10.1016/0024-3205(88)90094-x. [DOI] [PubMed] [Google Scholar]
  29. Nilsson-Ehle P., Belfrage P. A monoglyceride hydrolyzing enzyme in human postheparin plasma. Biochim Biophys Acta. 1972 May 23;270(1):60–64. doi: 10.1016/0005-2760(72)90177-4. [DOI] [PubMed] [Google Scholar]
  30. Patsch J. R., Prasad S., Gotto A. M., Jr, Bengtsson-Olivecrona G. Postprandial lipemia. A key for the conversion of high density lipoprotein2 into high density lipoprotein3 by hepatic lipase. J Clin Invest. 1984 Dec;74(6):2017–2023. doi: 10.1172/JCI111624. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Peters T., Jr Serum albumin. Adv Protein Chem. 1985;37:161–245. doi: 10.1016/s0065-3233(08)60065-0. [DOI] [PubMed] [Google Scholar]
  32. Poensgen J. Apolipoprotein C-1 inhibits the hydrolysis by phospholipase A2 of phospholipids in liposomes and cell membranes. Biochim Biophys Acta. 1990 Feb 6;1042(2):188–192. doi: 10.1016/0005-2760(90)90006-j. [DOI] [PubMed] [Google Scholar]
  33. SARDA L., DESNUELLE P. Action de la lipase pancréatique sur les esters en émulsion. Biochim Biophys Acta. 1958 Dec;30(3):513–521. doi: 10.1016/0006-3002(58)90097-0. [DOI] [PubMed] [Google Scholar]
  34. Schulthess G., Lipka G., Compassi S., Boffelli D., Weber F. E., Paltauf F., Hauser H. Absorption of monoacylglycerols by small intestinal brush border membrane. Biochemistry. 1994 Apr 19;33(15):4500–4508. doi: 10.1021/bi00181a009. [DOI] [PubMed] [Google Scholar]
  35. Segrest J. P., Jones M. K., De Loof H., Brouillette C. G., Venkatachalapathi Y. V., Anantharamaiah G. M. The amphipathic helix in the exchangeable apolipoproteins: a review of secondary structure and function. J Lipid Res. 1992 Feb;33(2):141–166. [PubMed] [Google Scholar]
  36. Serdarevich B., Carroll K. K. Synthesis and characterization of 1- and 2-monoglycerides of anteiso fatty acids. J Lipid Res. 1966 Mar;7(2):277–284. [PubMed] [Google Scholar]
  37. Somma-Delpero C., Meignen J. M., Jacquème B., Serment H., Boyer J. Increased monoester lipase activity in red blood cells in late pregnancy. Acta Haematol. 1985;73(2):120–121. doi: 10.1159/000206298. [DOI] [PubMed] [Google Scholar]
  38. Spector A. A., Soboroff J. M. Long chain fatty acid methyl ester hydrolase activity in mammalian cells. Lipids. 1972 Mar;7(3):186–190. doi: 10.1007/BF02533061. [DOI] [PubMed] [Google Scholar]
  39. Tornqvist H., Belfrage P. Purification and some properties of a monoacylglycerol-hydrolyzing enzyme of rat adipose tissue. J Biol Chem. 1976 Feb 10;251(3):813–819. [PubMed] [Google Scholar]
  40. Tornqvist H., Nilsson-Ehle P., Belfrage P. Enzymes catalyzing the hydrolysis of long-chain monoacyglycerols in rat adipose tissue. Biochim Biophys Acta. 1978 Sep 28;530(3):474–486. doi: 10.1016/0005-2760(78)90167-4. [DOI] [PubMed] [Google Scholar]
  41. Verger R. Enzyme kinetics of lipolysis. Methods Enzymol. 1980;64:340–392. doi: 10.1016/s0076-6879(80)64016-6. [DOI] [PubMed] [Google Scholar]
  42. Verine A., Boyer J. Lipases operative at the fat cell surface: attempt at an integrated approach. Cell Biochem Funct. 1987 Jul;5(3):175–181. doi: 10.1002/cbf.290050304. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES