The MARCKS family of phospholipid binding proteins: regulation of phospholipase D and other cellular components

Sundaram, Meenakshi; Cook, Harold W.; Byers, David M.

doi:10.1139/o03-087

Cited by 87 publications

(85 citation statements)

References 108 publications

(125 reference statements)

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“…from at least three independent experiments. sion, migration, secretion, and phagocytosis in a variety of cell types (22,27). However, MARCKS and MRP have not been associated with S1P-mediated barrier enhancement in EC.…”

Section: Discussionmentioning

confidence: 95%

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Quantitative Proteomics Analysis of Human Endothelial Cell Membrane Rafts

Guo

Singleton

Rowshan

et al. 2007

Molecular & Cellular Proteomics

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Endothelial cell barrier dysfunction results in the increased vascular permeability observed in inflammation, tumor metastasis, angiogenesis, and atherosclerosis. Sphingosine 1-phosphate (S1P), a biologically active phosphorylated lipid growth factor released from activated platelets, enhances the endothelial cell barrier integrity in vitro and in vivo. To begin to identify the molecular mechanisms mediating S1P induced endothelial barrier enhancement, quantitative proteomics analysis (iTRAQ TM ) was performed on membrane rafts isolated from human pulmonary artery endothelial cells in the absence or presence of S1P stimulation. Our results demonstrated that S1P mediates rapid and specific recruitment (1 M, 5 min) of myristoylated alanine-rich protein kinase C substrate (MARCKS) and MARCKS-related protein (MRP) to membrane rafts. Western blot experiments confirmed these findings with both MARCKS and MRP. Finally, small interfering RNA-mediated silencing of MARCKS or MRP or both attenuates S1P-mediated endothelial cell barrier enhancement. These data suggest the regulation of S1P-mediated endothelial cell barrier enhancement via the cell specific localization of MARCKS and MRP and validate the utility of proteomics approaches in the identification of novel molecular targets. Molecular & Cellular Proteomics 6:689 -696, 2007.The pulmonary endothelium serves as a semipermeable cellular barrier between blood and the interstitium and airspaces of the lung. Endothelial cell (EC) 1 barrier dysfunction results in increased vascular permeability, which is a cardinal feature of inflammation and an essential component of tumor metastasis, angiogenesis, and atherosclerosis. Proteins and lipids released after platelet activation have long been appreciated as enhancing the integrity of the microcirculation (1, 2). Sphingosine 1-phosphate (S1P), a biologically active phosphorylated lipid growth factor released from platelet, has multiple EC effects, including promoting barrier integrity in vivo and in vitro across both human and bovine pulmonary artery and lung microvascular ECs (3, 4). Our previous studies demonstrated S1P mediated barrier enhancement to be dependent on S1P binding to its major surface receptor, S1P 1 ; activation of the small GTPase, Rac1; and rearrangement of the cortical actin cytoskeleton (3, 5, 6). Very recently, we further detailed the essential involvement of PI 3-kinase, Tiam1, and ␣-actinin in specialized membrane domains (i.e. membrane rafts) in S1P-treated EC barrier regulation (7). However, the underlying signaling mechanisms by which S1P increases vascular integrity via signaling to the endothelial cytoskeleton remain poorly understood.As defined by the recent Keystone Symposium on lipid rafts and cell function (March 23-28, 2006, in Steamboat Springs, CO) (8), "Membrane rafts are small (10 -200 nm), heterogeneous, highly dynamic, sterol-and sphingolipid-enriched domains that compartmentalize cellular processes. Small rafts can sometimes be stabilized to form larger platforms through protein-protei...

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Section: Discussionmentioning

confidence: 95%

“…Both proteins contain three highly conserved domains including a myristoylated N terminus and an effector domain (22). The unusual biochemical properties and multiple interactions of MARCKS and MRP have led to a variety of proposed functions at the molecular level.…”

Section: Discussionmentioning

confidence: 99%

Quantitative Proteomics Analysis of Human Endothelial Cell Membrane Rafts

Guo

Singleton

Rowshan

et al. 2007

Molecular & Cellular Proteomics

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“…Second, PLD1 can be maintained inactive due to low availability of specific activators in rafts. It has been proposed that the myristoylated alanine-rich C kinase substrate (MARCKS), which resides in lipid rafts, diminishes the availability of PIP 2 due to its propensity to bind PIP 2 with a high affinity (38). It is noteworthy that MARCKS inhibits the PLC-catalyzed PIP 2 hydrolysis at physiological concentrations (39).…”

Section: Discussionmentioning

confidence: 99%

Disruption of Lipid Rafts Stimulates Phospholipase D Activity in Human Lymphocytes: Implication in the Regulation of Immune Function

Diaz

Mébarek-Azzam

Benzaria

et al. 2005

The Journal of Immunology

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Recent evidence suggests that phospholipase D (PLD) can be regulated through its association/dissociation to lipid rafts. We show here that modifying lipid rafts either by cholesterol depletion using methyl-β-cyclodextrin and filipin or by conversion of sphingomyelin to ceramide with exogenous bacterial sphingomyelinase (bSMase) markedly activated the PLD of human PBMC. bSMase was the most potent PLD activator, giving maximal 6- to 7-fold increase in PLD activity. Triton X-100-treated lysates prepared from control PBMC and from bSMase-treated cells were fractionated by centrifugation on sucrose density gradient. We observed that bSMase treatment of the cells induced a larger ceramide increase in raft than in nonraft membranes and displaced both the Src kinase Lck and PLD1 out of the raft fractions. In addition, the three raft-modifying agents markedly inhibited the lymphoproliferative response to mitogenic lectin. To examine further the potential role of PLD activation in the control of lymphocyte responses, we transiently overexpressed either of the PLD1 and PLD2 isoforms in Jurkat cells and analyzed the phorbol ester plus ionomycin-induced expression of IL-2 mRNA, which is one of the early responses of lymphocyte to activation. We observed a 43% decrease of IL-2 mRNA level in Jurkat cells overexpressing PLD1 as compared with mock- or PLD2-transfected cells, which indicates that elevated PLD1, but not PLD2, activity impairs lymphocyte activation. Altogether, the present results support the hypothesis that PLD1 is activated by exclusion from lipid rafts and that this activation conveys antiproliferative signals in lymphoid cells.

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“…ROCK can also act cooperatively with protein kinase C to induce MARCKS phosphorylation (67). LPS induces MARCKS phosphorylation (70), which inhibits its association with the plasma membrane and promotes cytosolic localization (71)(72)(73). LPS can also regulate NHE1 (74), a protein implicated in cell stress responses through regulation of intracellular pH and actin cytoskeletal dynamics (75,76).…”

Section: Discussionmentioning

confidence: 99%

CD44 Regulates Hepatocyte Growth Factor-mediated Vascular Integrity

Singleton

Salgia

Moreno‐Vinasco

et al. 2007

Journal of Biological Chemistry

111

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The preservation of vascular endothelial cell (EC) barrier integrity is critical to normal vessel homeostasis, with barrier dysfunction being a feature of inflammation, tumor angiogenesis, atherosclerosis, and acute lung injury. Therefore, agents that preserve or restore vascular integrity have important therapeutic implications. In this study, we explored the regulation of hepatocyte growth factor (HGF)-mediated enhancement of EC barrier function via CD44 isoforms. We observed that HGF promoted c-Met association with CD44v10 and recruitment of c-Met into caveolin-enriched microdomains (CEM) containing CD44s (standard form). Treatment of EC with CD44v10-blocking antibodies inhibited HGF-mediated c-Met phosphorylation and c-Met recruitment to CEM. Silencing CD44 expression (small interfering RNA) attenuated HGF-induced recruitment of c-Met, Tiam1 (a Rac1 exchange factor), cortactin (an actin cytoskeletal regulator), and dynamin 2 (a vesicular regulator) to CEM as well as HGF-induced trans-EC electrical resistance. In addition, silencing Tiam1 or dynamin 2 reduced HGF-induced Rac1 activation, cortactin recruitment to CEM, and EC barrier regulation. We observed that both HGF-and high molecular weight hyaluronan (CD44 ligand)-mediated protection from lipopolysaccharide-induced pulmonary vascular hyperpermeability was significantly reduced in CD44 knock-out mice, thus validating these in vitro findings in an in vivo murine model of inflammatory lung injury. Taken together, these results suggest that CD44 is an important regulator of HGF/c-Met-mediated in vitro and in vivo barrier enhancement, a process with essential involvement of Tiam1, Rac1, dynamin 2, and cortactin.

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The MARCKS family of phospholipid binding proteins: regulation of phospholipase D and other cellular components

Cited by 87 publications

References 108 publications

Quantitative Proteomics Analysis of Human Endothelial Cell Membrane Rafts

Quantitative Proteomics Analysis of Human Endothelial Cell Membrane Rafts

Disruption of Lipid Rafts Stimulates Phospholipase D Activity in Human Lymphocytes: Implication in the Regulation of Immune Function

CD44 Regulates Hepatocyte Growth Factor-mediated Vascular Integrity

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