Identification and characterization of a phospholipase C activity in resident mouse peritoneal macrophages. Inhibition of the enzyme by phenothiazines

Wightman, Paul D.; Dahlgren, Mary Ellen; Hall, James C.; Davies, P.L.; Bonney, Robert J.

doi:10.1042/bj1970523

Cited by 52 publications

(13 citation statements)

References 23 publications

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“…8) (54). This phospholipase is Ca2+-dependent (55,56), and the additional availability of intracellular free Ca2+ in LPS-primed neutrophils might enhance such activity. Enhanced PLA2 activity and release of arachidonic acid has been measured in neutrophils primed with granulocyte-macrophage colony-stimulating factor (57,58).…”

Section: Resultsmentioning

confidence: 99%

Lipopolysaccharide priming of human neutrophils for an enhanced respiratory burst. Role of intracellular free calcium.

Forehand¹,

Pabst²,

Phillips³

et al. 1989

J. Clin. Invest.

206

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Section: Resultsmentioning

confidence: 99%

Lipopolysaccharide priming of human neutrophils for an enhanced respiratory burst. Role of intracellular free calcium.

Forehand¹,

Pabst²,

Phillips³

et al. 1989

J. Clin. Invest.

206

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“…Later, Franson et al identified a phospholipase A 1 (PLA 1 ) and A 2 (PLA 2 ) in the soluble fraction of rat liver lysosomes [7]. Lysosomal phospholipase A activities were subsequently reported in a number of additional tissues and cell types including rat myocardial preparations [8], rabbit alveolar macrophages [9], arterial smooth muscle cells [10], rat testes [11], kidney cortex [12], and mouse peritoneal macrophages [13]. …”

Section: Introductionmentioning

confidence: 99%

“…The lysosomal PLA 1 was characterized by enhanced enzyme activity when the substrate was dispersed into inert detergent [14]. By contrast, the non-ionic detergent, Triton X-100, inhibited lysosomal PLA 2 activity [13], suggesting that the enzyme requires a lipid bilayer structure to catalyze the substrate degradation.…”

Section: Introductionmentioning

confidence: 99%

Group XV phospholipase A2, a lysosomal phospholipase A2

Shayman

Kelly

Kollmeyer

et al. 2011

Progress in Lipid Research

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A phospholipase A 2 was identified from MDCK cell homogenates with broad specificity toward glycerophospholipids including phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and phosphatidylglycerol. The phospholipase has the unique ability to transacylate short chain ceramides. This phospholipase is calcium-independent, localized to lysosomes, and has an acidic pH optimum. The enzyme was purified from bovine brain and found to be a water-soluble glycoprotein consisting of a single peptide chain with a molecular weight of 45 kDa. The primary structure deduced from the DNA sequences is highly conserved between chordates. The enzyme was named lysosomal phospholipase A2 (LPLA 2 ) and subsequently designated group XV phospholipase A 2 . LPLA 2 has 49 percent of amino acid sequence identity to lecithin cholesterol acyltransferase and is a member of the αβ-hydrolase superfamily. LPLA 2 is highly expressed in alveolar macrophages. A marked accumulation of glycerophospholipids and extensive lamellar inclusion bodies, a hallmark of cellular phospholipidosis, is observed in alveolar macrophages in LPLA 2 −/− mice. This defect can also be reproduced in macrophages that are exposed to cationic amphiphilic drugs such as amiodarone. In addition, older LPLA 2 −/− mice develop a phenotype similar to human autoimmune disease. These observations indicate that LPLA 2 may play a primary role in phospholipid homeostasis, drug toxicity, and host defense.

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“…Using ap63 as a substrate, we found that the B. cereus lipase activity was inhibited by 5 (24), it may be that if the membrane anchor of p63 is cleaved at all, it is cleaved by a macrophage enzyme. In the African trypanosomes, VSGs are shed when bloodstream trypanosomes differentiate to a form lacking a surface coat (25).…”

Section: Resultsmentioning

confidence: 97%

Leishmania and Trypanosoma surface glycoproteins have a common glycophospholipid membrane anchor.

Bordier¹,

Etges²,

Ward³

et al. 1986

Proc. Natl. Acad. Sci. U.S.A.

108

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The variant surface glycoprotein (VSG) ofthe African trypanosomes is the major membrane protein of the plasma membrane of the bloodstream stage of the parasite. It is anchored in the plasma membrane by a glycolipid covalently bound to the C-terminal amino acid of the protein. The by phase-separation in Triton X-114 (TX-114) and chromatography on DEAE-cellulose and Mono Q (Pharmacia) were performed as described (7). Phospholipase from T. brucei ILTat 1.25 was obtained by solubilization of VSG-depleted membranes in n-octyl glucoside, followed by covalent chromatography on Affi-Gel 501 (Bio-Rad), or by TX-114 extraction from VSG-depleted membranes, followed by phaseseparation at 37°C (J.W. and C.B., unpublished method). Purified MITat 1.6 mfVSG was radioiodinated on an affinity column of antibody as described (14), and ap63 was radioiodinated on living L. major promastigotes and then purified to homogeneity (3, 7). Phospholipase C, type III, from Bacillus cereus was obtained from Sigma. Antibody to the CRD of T. brucei was prepared and affinity-purified as described (5, 9). Staphylococcus aureus protein A conjugated to horseradish peroxidase was from Miles-Yeda (Rehovot, Israel). Endoglycosidase F was a gift from J. Kaufman (Basel Institute of Immunology).Gel Electrophoresis. NaDodSO4/polyacrylamide gel electrophoresis was done according to the method of Laemmli (15). Gels were fixed and stained with 0.1% Coomassie blue or were prepared for 3H fluorography by immersion in Amplify (Amersham). For immunoblotting, samples were fractionated by gel electrophoresis and were electrophoretically transferred to nitrocellulose paper, which then was stained with Ponceau S and photographed. After destaining, the nitrocellulose was saturated with 1% gelatin in TBS (10 mM Tris HCl/150 mM NaCl, pH 7.4) and was then incubated overnight with anti-CRD (2 ,tg/ml) prior to detection with S.aureus protein A-horseradish peroxidase.Phospholipase Digestions. Removal of [3H]myristate from ap63 was monitored by incubation of 2 ,ug of the labeled ap63 in 160 ,ul of TBS with 0.5 mM dithiothreitol and 0.05% Triton X-100 (TX-100) for 1 hr at 30°C in the presence (0.45 ,ul/ml) or absence of affinity-purified lipase. After digestion, samples were heated and reduced prior to analysis by electrophoresis in a 7.5-15% polyacrylamide gradient gel. To monitor conversion of the amphiphilic forms of p63 and VSG Abbreviations: TX-114, Triton X-114; TX-100, Triton X-100; VSG, variant surface glycoprotein; mfVSG and sVSG, membrane-form and soluble VSG, respectively; CRD, crossreacting determinant of VSG; ap63 and hp63, aipphiphilic and hydrophilic forms, respectively, of the p63 antigen of Leishmania. 5988The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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Identification and characterization of a phospholipase C activity in resident mouse peritoneal macrophages. Inhibition of the enzyme by phenothiazines

Cited by 52 publications

References 23 publications

Lipopolysaccharide priming of human neutrophils for an enhanced respiratory burst. Role of intracellular free calcium.

Lipopolysaccharide priming of human neutrophils for an enhanced respiratory burst. Role of intracellular free calcium.

Group XV phospholipase A2, a lysosomal phospholipase A2

Leishmania and Trypanosoma surface glycoproteins have a common glycophospholipid membrane anchor.

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