Comparative proteomic analysis of myotube <b><i>caveolae</i></b> after milli‐calpain deregulation

Goudenège, Sébastien; Dargelos, Elise; Claverol, Stéphane; Bonneu, Marc; Cottin, Patrick; Poussard, Sylvie

doi:10.1002/pmic.200700124

Cited by 15 publications

(12 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Caveolar-raft localization of calpains has been described in myotubes (40), Jurkat T-cells (41), neutrophils subjected to chemotactic stimulation (42). The silencing of calpain small subunit-1 by siRNA resulted into capillary morphogenesis impairment, in agreement with the indication that calpain mediates VEGF-induced angiogenic effects by a PKC and Rho GTPase.…”

Section: Fig 3 Quantitative Rt-pcr and Western Blotting Analisys Ofsupporting

confidence: 67%

Proteomic Identification of VEGF-dependent Protein Enrichment to Membrane Caveolar-raft Microdomains in Endothelial Progenitor Cells

Chillà

Magherini

Margheri

et al. 2013

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

Endothelial cell caveolar-rafts are considered functional platforms that recruit several pro-angiogenic molecules to realize an efficient angiogenic program. Here we studied the differential caveolar-raft protein composition of endothelial colony-forming cells following stimulation with VEGF, which localizes in caveolae on interaction with its type-2 receptor. Endothelial colony-forming cells are a cell population identified in human umbilical blood that show all the properties of an endothelial progenitor cell and a high proliferative rate. Two-dimensional gel electrophoresis analysis was coupled with mass spectrometry to identify candidate proteins. The twenty-eight differentially expressed protein spots were grouped according to their function using Gene Ontology classification. In particular, functional categories relative to cell death inhibition and hydrogen peroxide metabolic processes resulted enriched. In these categories, Peroxiredoxin-2 and 6, that control hydrogen peroxide metabolic processes, are the main enriched molecules together with the anti-apoptotic 78 kDa glucose regulated protein. Some of the proteins we identified had never before identified as caveolar-raft components. Other identified proteins include calpain small subunit-1, known to mediates angiogenic response to VEGF, gelsolin, which regulates stress fiber assembly, and annexin A3, an angiogenic mediator that induces VEGF production. We validated the functional activity of the above proteins, showing that the siRNA silencing of these resulted in the inhibition of capillary morphogenesis. Overall, our data show that VEGF stimulation triggers the caveolar-raft recruitment of proteins that warrant a physiological amount of reactive oxygen species to maintain a proper angiogenic function of endothelial colony-forming cells and preserve the integrity of the actin cytoskeleton. Molecular & Cellular

show abstract

Section: Fig 3 Quantitative Rt-pcr and Western Blotting Analisys Ofsupporting

confidence: 67%

Proteomic Identification of VEGF-dependent Protein Enrichment to Membrane Caveolar-raft Microdomains in Endothelial Progenitor Cells

Chillà

Magherini

Margheri

et al. 2013

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

show abstract

“…These changes may reflect a change in compartmentalization of TRPM7 as suggested previously (22) and are supported here where we show that TRPM7 interacts with caveolae/lipid rafts in activated cells, as shown by immunofluorescence and by localization in flotillin-2-containing cholesterol-rich cell fractions. TRPM7/caveolae/lipid raft association may facilitate TRPM7 scaffolding to cell membrane receptors, such as G protein-coupled receptors and to signaling molecules, such as calpain, which has been shown to be a caveolae-associated protein (15,37).…”

Section: Discussionmentioning

confidence: 99%

Bradykinin regulates calpain and proinflammatory signaling through TRPM7-sensitive pathways in vascular smooth muscle cells

Yogi¹,

Callera²,

Tostes³

et al. 2009

American Journal of Physiology-Regulatory, Integrative and Comp

View full text Add to dashboard Cite

Yogi A, Callera GE, Tostes R, Touyz RM. Bradykinin regulates calpain and proinflammatory signaling through TRPM7-sensitive pathways in vascular smooth muscle cells. Am J Physiol Regul Integr Comp Physiol 296: R201-R207, 2009. First published September 17, 2008 doi:10.1152/ajpregu.90602.2008.-Transient receptor potential melastatin-7 (TRPM7) channels have recently been identified to be regulated by vasoactive agents acting through G protein-coupled receptors in vascular smooth muscle cells (VSMC). However, downstream targets and functional responses remain unclear. We investigated the subcellular localization of TRPM7 in VSMCs and questioned the role of TRPM7 in proinflammatory signaling by bradykinin. VSMCs from Wistar-Kyoto rats were studied. Cell fractionation by sucrose gradient and differential centrifugation demonstrated that in bradykinin-stimulated cells, TRPM7 localized in fractions corresponding to caveolae. Immunofluorescence confocal microscopy revealed that TRPM7 distributes along the cell membrane, that it has a reticular-type intracellular distribution, and that it colocalizes with flotillin-2, a marker of lipid rafts. Bradykinin increased expression of calpain, a TRPM7 target, and stimulated its cytosol/membrane translocation, an effect blocked by 2-APB (TRPM7 inhibitor) and U-73122 (phospholipase C inhibitor), but not by chelerythrine (PKC inhibitor). Expression of proinflammatory mediators VCAM-1 and cyclooxygenase-2 (COX-2) was time-dependently increased by bradykinin. This effect was blocked by Hoe-140 (B 2 receptor blocker) and 2-APB. Our data demonstrate that in bradykinin-stimulated VSMCs: 1) TRPM7 is upregulated, 2) TRPM7 associates with cholesterol-rich microdomains, and 3) calpain and proinflammatory mediators VCAM-1 and COX2 are regulated, in part, via TRPM7-and phospholipase C-dependent pathways through B 2 receptors. These findings identify a novel signaling pathway for bradykinin, which involves TRPM7. Such phenomena may play a role in bradykinin/B 2 receptor-mediated inflammatory responses in vascular cells.

show abstract

“…Although the association between cytoskeletal components and MLR/caveolae had been previously described[15, 16], recent evidence has extended the notion that cytoskeletal components (e.g., actin, tubulin, vinculin, filamin, and tau)[17, 18] can localize to MLR and be platforms for cytoskeletal tethering and for communication to the extracellular matrix (ECM) via integrins, cadherins, occludins, and other cellular adhesion molecules (CAMs). Moreover, MLR can cluster and this clustering may depend upon cholesterol and actin tethering to the membrane[19].…”

Section: Introductionmentioning

confidence: 99%

Interaction of membrane/lipid rafts with the cytoskeleton: Impact on signaling and function

Head

Patel

Insel

2014

Biochimica et Biophysica Acta (BBA) - Biomembranes

422

186

View full text Add to dashboard Cite

Summary The plasma membrane in eukaryotic cells contains microdomains that are enriched in certain glycosphingolipids, gangliosides, and sterols (such as cholesterol) to form membrane/lipid rafts (MLR). These regions exist as caveolae, morphologically observable flask-like invaginations, or as a less easily detectable planar form. MLR are scaffolds for many molecular entities, including signaling receptors and ion channels that communicate extracellular stimuli to the intracellular milieu. Much evidence indicates that this organization and/or the clustering of MLR into more active signaling platforms depends upon interactions with and dynamic rearrangement of the cytoskeleton. Several cytoskeletal components and binding partners, as well as enzymes that regulate the cytoskeleton, localize to MLR and help regulate lateral diffusion of membrane proteins and lipids in response to extracellular events (e.g., receptor activation, shear stress, electrical conductance, and nutrient demand). MLR regulate cellular polarity, adherence to the extracellular matrix, signaling events (including ones that affect growth and migration), and are sites of cellular entry of certain pathogens, toxins and nanoparticles. The dynamic interaction between MLR and the underlying cytoskeleton thus regulates many facets of the function of eukaryotic cells and their adaptation to changing environments. Here, we review general features of MLR and caveolae and their role in several aspects of cellular function, including polarity of endothelial and epithelial cells, cell migration, mechanotransduction, lymphocyte activation, neuronal growth and signaling, and a variety of disease settings.

show abstract

Comparative proteomic analysis of myotube caveolae after milli‐calpain deregulation

Cited by 15 publications

References 35 publications

Proteomic Identification of VEGF-dependent Protein Enrichment to Membrane Caveolar-raft Microdomains in Endothelial Progenitor Cells

Proteomic Identification of VEGF-dependent Protein Enrichment to Membrane Caveolar-raft Microdomains in Endothelial Progenitor Cells

Bradykinin regulates calpain and proinflammatory signaling through TRPM7-sensitive pathways in vascular smooth muscle cells

Interaction of membrane/lipid rafts with the cytoskeleton: Impact on signaling and function

Contact Info

Product

Resources

About