The Effect of Sn-2 Fatty Acid Substitution on Phospholipase C Enzyme Activities

Overview

Journal Biochem J

Specialty Biochemistry

Date 1987 Jun 15

PMID 3446171

Citations 3

Authors

J S Bomalaski

M A Clark

Affiliations

Soon will be listed here.

Abstract

The human monocyte cell line U937 expresses phospholipase A2 and phospholipase C activities and produces eicosanoids. The phospholipase C (PLC) activity exhibits substrate preference for phosphatidyl-choline (PC), rather than phosphatidylinositol or phosphatidylethanolamine. In order to characterize the PLC activity found in these cells, the effects of substitution of the sn-2 fatty acid on this activity were examined. PC substrates with palmitic acid (PC-2P), oleic acid (PC-2O), arachidonic acid (PC-2A) and linoleic acid (PC-2L) at the sn-2 position were used. The sn-1 fatty acid was palmitic acid. PC-2L and PC-2A with the longer-chain less-saturated fatty acids linoleic acid and arachidonic acid esterified at sn-2 were found to be better substrates for PLC activity than PC-2P or PC-2O in these cells. This preference was maintained even when substrate phospholipid was solubilized in non-ionic, anionic, cationic and zwitterionic amphiphiles. Furthermore, when a 500-fold excess of 1,2-diolein or 1,2-dipalmitin was added to the reaction, the specificity of the PLC activity for PC-2A and PC-2L remained unchanged. When similar experiments were performed with phosphatidylinositol as a substrate, we did not observe any effect when the sn-2 position was altered. These data show that the fatty acid constituent at the sn-2 position affects the observed PLC activity when phosphatidylcholine, but not phosphatidylinositol, is used as a substrate by these cells.

Citing Articles

Acyl chain and head group regulation of phospholipid catabolism in senescing carnation flowers.

Brown J, Chambers J, Thompson J Plant Physiol. 1991; 95(3):909-16.

PMID: 16668071 PMC: 1077623. DOI: 10.1104/pp.95.3.909.

Is metabolism an important arbiter of anticancer activity of ether lipids? Metabolism of SRI 62-834 and hexadecylphosphocholine by [31P]-NMR spectroscopy and comparison of their cytotoxicities with those of their metabolites.

Bishop F, Dive C, Freeman S, Gescher A Cancer Chemother Pharmacol. 1992; 31(2):85-92.

PMID: 1451237 DOI: 10.1007/BF00685092.

Effects of polyunsaturated fatty acids on interleukin-2-dependent T cell growth.

Borofsky M, Zurier R, Rosenbaum H, Weiner D, Williams W Immunol Res. 1992; 11(2):154-64.

PMID: 1431424 DOI: 10.1007/BF02918620.

References

Low M, Weglicki W . Resolution of myocardial phospholipase C into several forms with distinct properties. Biochem J. 1983; 215(2):325-34. PMC: 1152400. DOI: 10.1042/bj2150325. View

Pawlowski N, Kaplan G, Hamill A, COHN Z, Scott W . Arachidonic acid metabolism by human monocytes. Studies with platelet-depleted cultures. J Exp Med. 1983; 158(2):393-412. PMC: 2187343. DOI: 10.1084/jem.158.2.393. View

Lenstra R, Mauco G, Chap H, DOUSTE-BLAZY L . Studies on enzymes related to diacylglycerol production in activated platelets. I. Phosphatidylinositol-specific phospholipase C: further characterization using a simple method for determination of activity. Biochim Biophys Acta. 1984; 792(2):199-206. DOI: 10.1016/0005-2760(84)90223-6. View

Eisen D, Bartolf M, Franson R . Inhibition of lysosomal phospholipases C and A in rabbit alveolar macrophages, polymorphonuclear leukocytes and rat liver by sodium bisulfite. Biochim Biophys Acta. 1984; 793(1):10-7. DOI: 10.1016/0005-2760(84)90047-x. View

Low M, Carroll R, Weglicki W . Multiple forms of phosphoinositide-specific phospholipase C of different relative molecular masses in animal tissues. Evidence for modification of the platelet enzyme by Ca2+-dependent proteinase. Biochem J. 1984; 221(3):813-20. PMC: 1144111. DOI: 10.1042/bj2210813. View

Holub B, Celi B . Evaluation of the fatty acid selectivity of a phosphatidylinositol-specific cytosolic phospholipase C from pig and human platelets. Can J Biochem Cell Biol. 1984; 62(2-3):115-20. DOI: 10.1139/o84-017. View

Ross M, Deems R, Jesaitis A, Dennis E, Ulevitch R . Phospholipase activities of the P388D1 macrophage-like cell line. Arch Biochem Biophys. 1985; 238(1):247-58. DOI: 10.1016/0003-9861(85)90162-6. View

Wolf R, Gross R . Identification of neutral active phospholipase C which hydrolyzes choline glycerophospholipids and plasmalogen selective phospholipase A2 in canine myocardium. J Biol Chem. 1985; 260(12):7295-303. View

Roberts M . Charged detergents enhance the activity of phospholipase C (Bacillus cereus) towards micellar short-chain phosphatidylcholine. Biochim Biophys Acta. 1985; 831(1):133-41. DOI: 10.1016/0167-4838(85)90160-8. View

10.

Bomalaski J, Clark M, Douglas S, Zurier R . Enhanced phospholipase A2 and C activities of peripheral blood polymorphonuclear leukocytes from patients with rheumatoid arthritis. J Leukoc Biol. 1985; 38(5):649-54. DOI: 10.1002/jlb.38.5.649. View

11.

DeBose C, Coury L, Roberts M . Sensitivity of phospholipase C (Bacillus cereus) activity to phosphatidylcholine structural modifications. Biochim Biophys Acta. 1985; 837(3):325-35. DOI: 10.1016/0005-2760(85)90056-6. View

12.

Waite M . Approaches to the study of mammalian cellular phospholipases. J Lipid Res. 1985; 26(12):1379-88. View

13.

Bomalaski J, Clark M, Zurier R . Enhanced phospholipase activity in peripheral blood monocytes from patients with rheumatoid arthritis. Arthritis Rheum. 1986; 29(3):312-8. DOI: 10.1002/art.1780290302. View

14.

Bomalaski J, Goldstein C, Dailey A, Douglas S, Zurier R . Uptake of fatty acids and their mobilization from phospholipids in cultured monocyte-macrophages from rheumatoid arthritis patients. Clin Immunol Immunopathol. 1986; 39(2):198-212. DOI: 10.1016/0090-1229(86)90084-x. View

15.

Bomalaski J, Hirata F, Clark M . Aspirin inhibits phospholipase C. Biochem Biophys Res Commun. 1986; 139(1):115-21. DOI: 10.1016/s0006-291x(86)80087-0. View

16.

Flower R . Drugs which inhibit prostaglandin biosynthesis. Pharmacol Rev. 1974; 26(1):33-67. View

17.

Helenius A, Simons K . Solubilization of membranes by detergents. Biochim Biophys Acta. 1975; 415(1):29-79. DOI: 10.1016/0304-4157(75)90016-7. View

18.

Bradford M . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72:248-54. DOI: 10.1016/0003-2697(76)90527-3. View

19.

Little C . Phospholipase C from Bacillus cereus. Action on some artificial lecithins. Acta Chem Scand B. 1977; 31(4):267-72. DOI: 10.3891/acta.chem.scand.31b-0267. View

20.

Otnaess A, Little C, Sletten K, Wallin R, Johnsen S, Flengsrud R . Some characteristics of phospholipase C from Bacillus cereus. Eur J Biochem. 1977; 79(2):459-68. DOI: 10.1111/j.1432-1033.1977.tb11828.x. View