Inhibition of Cytosolic Phospholipase A2 by Annexin V in Differentiated Permeabilized HL-60 Cells

Mira, Jean‐Paul; Dubois, Thierry; Oudinet, Jean–Paul; Lukowski, Sandra; Russo‐Marie, Françoise; Geny, Blandine

doi:10.1074/jbc.272.16.10474

Cited by 107 publications

(52 citation statements)

References 68 publications

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“…Such proteins have already been proposed to explain the interaction of annexin with cell membranes at low Ca# + concentrations [29] and might provide an explanation for the ability of annexin V to inhibit cPLA # activity in permeabilized HL-60 cells [30].…”

Section: Discussionmentioning

confidence: 97%

Inhibition of human cytosolic phospholipase A2 by human annexin V

Buckland

Wilton

1998

Biochemical Journal

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The ability of annexins, particularly annexin 1 (lipocortin 1), to inhibit phospholipase A2 (PLA2) is well known and a substrate depletion mechanism is now widely accepted as the explanation for most inhibitory studies. However, there are only a very limited number of reported studies involving annexins and the high-molecular-mass cytosolic PLA2 (cPLA2). In this study we have examined the effect of human recombinant annexin V, a potentially abundant cytosolic protein, on the ability of human recombinant cPLA2 to hydrolyse a variety of phospholipid substrates. The results show clearly that, under the conditions of our study, annexin V can inhibit cPLA2 activity by a mechanism of substrate depletion and that this inhibition is dependent on the nature of the phospholipids and the concentration of Ca2+ ions in the assay. The hydrolysis of 1-stearoyl 2-arachidonyl phosphatidylcholine by cPLA2 was not significantly affected by annexin V over a range of Ca2+ concentrations (1 μM-2.5 mM), a result that presumably reflects the zwitterionic nature of the phospholipid and the known inability of annexins to bind to such interfaces. In contrast, the hydrolysis of dioleoyl phosphatidylglycerol, which is an effective anionic phospholipid substrate for this enzyme, and more significantly that of 1-stearoyl 2-arachidonyl phosphatidic acid, were readily inhibited by annexin V, although these effects were Ca2+-dependent. The Ca2+ concentrations required for inhibition in the assay system in vitro are greater than those associated with Ca2+-stimulated events within the cell, suggesting that a role for annexin V in regulating cPLA2 activity might not involve a substrate depletion mechanism in vivo unless factors in addition to Ca2+ and phospholipids contribute to the binding of annexin V to cell membranes.

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Section: Discussionmentioning

confidence: 97%

Inhibition of human cytosolic phospholipase A2 by human annexin V

Buckland

Wilton

1998

Biochemical Journal

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“…The M1234 cDNA was recloned to remove the 5Ј-base pairs responsible for the three extra N-terminal amino acids after expression (21). The annexin A5 mutant M23 was generated by subcloning the mutations D144N and E228A into the annexin A5 cDNA.…”

Section: Methodsmentioning

confidence: 99%

“…We hypothesized that the trimers bend the membrane and provide the driving force for the reversion of membrane movement. To test the hypothesis we generated the annexin A5 mutant M23 (21), which by prediction from the available structural data (19,27) binds to PtdSer but lacks the ability to form trimers. Using a novel FRET assay we showed that annexin A5 but not M23 develops FRET when bound to a phospholipid surface (Fig.…”

Section: A Novel Pinocytic Pathway Internalizes Annexin A5 Into Cellsmentioning

confidence: 99%

Cell Surface-expressed Phosphatidylserine and Annexin A5 Open a Novel Portal of Cell Entry

Kenis¹,

Genderen²,

Bennaghmouch³

et al. 2004

Journal of Biological Chemistry

142

139

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Expression of phosphatidylserine (PtdSer) at the cell surface is part of the membrane dynamics of apoptosis. Expressed phosphatidylserine functions as an "eat me" flag toward phagocytes. Here, we report that the expressed phosphatidylserine forms part of a hitherto undescribed pinocytic pathway. Annexin A5, a phosphatidylserine-binding protein, binds to and polymerizes through protein-protein interactions on membrane patches expressing phosphatidylserine. The two-dimensional protein network of annexin A5 at the surface prevents apoptotic body formation without interfering with the progression of apoptosis as demonstrated by activation of caspase-3, PtdSer exposure, and DNA fragmentation. The annexin A5 protein network bends the membrane patch nanomechanically into the cell and elicits budding, endocytic vesicle formation, and cytoskeleton-dependent trafficking of the endocytic vesicle. Annexin A1, which binds to PtdSer without forming a two-dimensional protein network, does not induce the formation of endocytic vesicles. This novel pinocytic pathway differs from macropinocytosis, which is preceded by membrane ruffling and actin polymerization. We clearly showed that actin polymerization is not involved in budding and endocytic vesicle formation but is required for intracellular trafficking. The phosphatidylserine-annexin A5-mediated pinocytic pathway is not restricted to cells in apoptosis. We demonstrated that living tumor cells can take up substances through this novel portal of cell entry. This opens new avenues for targeted drug delivery and cell entry.

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“…The Cterminal region of annexin I binds to the C2 domain of cPLA 2 a and inhibits PLA 2 activity by specific interaction, 75) whereas annexin V inhibits cPLA 2 a activity mainly by substrate depletion. 76) Annexins II and III do not have any inhibitory effects. It is of interest that some annexins are upregulated by antiinflammatory glucocorticoids.…”

Section: -5 Regulation By Protein Interaction and Cleavagementioning

confidence: 99%

Regulatory Mechanism and Physiological Role of Cytosolic Phospholipase A2

Hirabayashi

Murayama

Shimizu

2004

Biological & Pharmaceutical Bulletin

214

184

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Cytosolic phospholipase A 2 a a (cPLA 2 a a) preferentially hydrolyzes phospholipids containing arachidonic acid and plays a key role in the biosynthesis of eicosanoids. This review discusses the essential features of cPLA 2 a a regulation and addresses new insights into the functional properties of this enzyme. Full activation of the enzyme requires Ca 2؉ binding to an N-terminal C2 domain and phosphorylation on serine residues. Ca 2؉ binding induces translocation of cPLA 2 a a from the cytosol to the perinuclear membranes. Serine phosphorylation is mediated by mitogen-activated protein kinases (MAPKs), Ca 2؉ /calmodulin-dependent protein kinase II, and MAPK-interacting kinase Mnk1. Interaction with proteins and lipids, which include vimentin, annexins, NADPH oxidase, phosphatidylcholine, phosphatidylinositol 4,5-bisphosphate (PIP 2 ), and ceramide-1-phosphate, can also modulate the activity of cPLA 2 a a. Recent evidence has established the physiological and pathological roles of cPLA 2 a a using cPLA 2 a a knockout mice. This enzyme has been implicated in fertility, striated muscle growth, renal concentration, postischemic brain injury, arthritis, inflammatory bone resorption, intestinal polyposis, pulmonary fibrosis, acute respiratory distress syndrome, and autoimmune encephalomyelitis. Now novel three paralogs, cPLA 2 b b, cPLA 2 g g, and cPLA 2 d d, have been identified in humans. cPLA 2 g g is distinct from others in that it is farnesylated and lacks the C2 domain. Biological roles for these new enzymes have not yet been defined.

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Inhibition of Cytosolic Phospholipase A2 by Annexin V in Differentiated Permeabilized HL-60 Cells

Cited by 107 publications

References 68 publications

Inhibition of human cytosolic phospholipase A2 by human annexin V

Inhibition of human cytosolic phospholipase A2 by human annexin V

Cell Surface-expressed Phosphatidylserine and Annexin A5 Open a Novel Portal of Cell Entry

Regulatory Mechanism and Physiological Role of Cytosolic Phospholipase A2

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