Palmitoylation at Cys595 is essential for PECAM-1 localisation into membrane microdomains and for efficient PECAM-1-mediated cytoprotection

Caroline T. Sardjono; Stacey N. Harbour; Jana C. Yip; Cathy Paddock; Susheela Tridandapani; Peter J. Newman; Denise E. Jackson

doi:10.1160/TH06-08-0459

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2006; 96(06): 756-766
DOI: 10.1160/TH06-08-0459

Platelets and Blood Cells

Schattauer GmbH

Palmitoylation at Cys⁵⁹⁵ is essential for PECAM-1 localisation into membrane microdomains and for efficient PECAM-1-mediated cytoprotection

Caroline T. Sardjono

¹Kronheimer Building, Burnet Institute incorporating the Austin Research Institute, Heidelberg, Victoria, Australia

,

Stacey N. Harbour

¹Kronheimer Building, Burnet Institute incorporating the Austin Research Institute, Heidelberg, Victoria, Australia

,

Jana C. Yip

¹Kronheimer Building, Burnet Institute incorporating the Austin Research Institute, Heidelberg, Victoria, Australia

,

Cathy Paddock

³Blood Research Institute, Blood Center of Wisconsin, Milwaukee, Wisconsin, USA

,

Susheela Tridandapani

²Department of Internal Medicine, The Ohio State University, Columbus, Ohio, USA

,

Peter J. Newman

³Blood Research Institute, Blood Center of Wisconsin, Milwaukee, Wisconsin, USA

,

Denise E. Jackson

¹Kronheimer Building, Burnet Institute incorporating the Austin Research Institute, Heidelberg, Victoria, Australia

› Institutsangaben

Abstract

Summary

The Ig-ITIM superfamily member, PECAM-1 acts as a negative regulator of ITAM-signalling pathways in platelets involving GPVI/FcR gamma chain and FcγRIIa. This negative feedback loop involves regulation of collagen and GPVI-dependent aggregation events, platelet-thrombus-growth on immobilised collagen under flow and FcγRIIa-mediated platelet responses. In this study, we show that PECAM-1 is selectively palmitoylated involving a thioester linkage with an unpaired cysteine residue at amino acid position 595 in its cytoplasmic domain. As palmitoylation is known to target proteins to membrane microdomains, we investigated the microdomain localisation for PECAM-1 in platelets and nucleated cells. In unstimulated platelets, ∼20% of PECAM-1 is localised toTriton-insoluble microdomain fractions and it does not increase with platelet activation by collagen, collagen-related peptide, thrombin-or human-aggregated IgG. PECAM-1 is in close physical proximity with GPVI in platelet microdomains. Removal of platelet cytoskeleton prior to sucrose-density-gradient separation showed that PECAM-1 was associated with both theTriton-soluble and membrane skeleton in microdomain-associated fractions. Disruption of microdomains by membrane-cholesterol depletion resulted in loss of PECAM-1 localisation to membrane microdomains. Mutational analysis of juxtamembrane cysteine residue to alanine (C595A) of human PECAM-1 resulted in loss of palmitoylation and a sixfold decrease in association with membrane microdomains. Functionally, the palmitoylated cysteine 595 residue is required, in part, for efficient PECAM-1-mediated cytoprotection. These results show that cysteine 595 is required for constitutive association of PECAM-1 with membrane microdomains and PECAM-1-mediated cytoprotection, where it may act as a crucial regulator of signaling and apoptosis events.

Keywords

Adhesion receptors - signal transduction - platelet physiology - lipid rafts - GP VI

Volltext

Referenzen

References
1 Simons K, Toomre D. Lipid rafts and signal transduction. Nature Rev Mol Cell Biol 2000; 01: 31-41.
2 Hua CT, Gamble JR, Vadas MA. et al. Recruitment and activation of SHP-1 protein-tyrosine phosphatase by human platelet endothelial cell adhesion molecule-1 (PECAM-1). Identification of immunoreceptor tyrosine-based inhibitory motif-like binding motifs and substrates. J Biol Chem 1998; 273: 28332-40.
3 Newman PJ. Switched at birth: a new family for PECAM-1. J Clin Invest 1999; 103: 5-9.
4 Jackson DE. The unfolding tale of PECAM-1. FEBS Lett 2004; 540: 7-14.
5 Muller WA, Weigl SA, Deng X. et al. PECAM-1 is required for transendothelial migration of leukocytes. J Exp Med 1993; 178: 449-60.
6 Mamdouh Z, Chen X, Pierini LM. et al. Targeting recycling of PECAM-1 from endothelial surface-connected compartments during diapedesis. Nature 2003; 421: 748-55.
7 Schimmenti LA, Yan HC, Madri JA. et al. Platelet endothelial cell adhesion molecule, PECAM-1, modulates cell migration. J Cell Physiol 1992; 153: 417-28.
8 Kim CS, Wang T, Madri JA. Platelet endothelial cell adhesion molecule-1 expression modulates endothelial cell migration in vitro . Lab Invest 1998; 78: 583-90.
9 Albelda SM, Lau KC, Chien P. et al. Role for platelet endothelial cell adhesion molecule-1 in macrophage Fc gamma receptor function. Am J Respir Cell Mol Biol 2004; 31: 246-55.
10 Brown S, Heinisch I, Ross E. et al. Apoptosis disables CD31-mediated cell detachment from phagocytes promoting binding and engulfment. Nature 2002; 418: 200-3.
11 DeLisser HM, Christofidou-Solomidou M, Strieter RM. et al. Involvement of endothelial PECAM-1/CD31 in angiogenesis. Am J Pathol 1997; 151: 671-7.
12 Gao C, Sun W, Christofidou-Solomidou M. et al. PECAM-1 functions as a specific and potent inhibitor of mitochondrial-dependent apoptosis. Blood 2003; 102: 169-79.
13 Wilkinson R, Lyons AB, Roberts D. et al. Platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31) acts as a regulator of B-cell development, B-cell antigen receptor (BCR)-mediated activation, and autoimmune disease. Blood 2002; 100: 184-93.
14 Newton-Nash DK, Newman PJ. A new role for platelet-endothelial cell adhesion molecule-1 (CD31): inhibition of TCR-mediated signal transduction. J Immunol 1999; 163: 682-8.
15 Wong MX, Roberts D, Bartley PA. et al. Absence of platelet endothelial cell adhesion molecule-1 (CD31) leads to increased severity of local and systemic IgE-mediated anaphylaxis and modulation of mast cell activation. J Immunol 2002; 168: 6455-62.
16 Jones KL, Hughan SC, Dopheide SM. et al. Platelet endothelial cell adhesion molecule-1 is a negative regulator of platelet-collagen interactions. Blood 2001; 98: 1456-63.
17 Patil S, Newman DK, Newman PJ. Platelet endothelial cell adhesion molecule-1 serves as an inhibitory receptor that modulates platelet responses to collagen. Blood 2001; 97: 1727-32.
18 Falati S, Patil S, Gross PL. et al. Platelet PECAM-1 inhibits thrombus formation in vivo . Blood 2006; 107: 535-41.
19 Thai le M, Ashman LK, Harbour SN. et al. Physical proximity and functional interplay of PECAM-1 with the Fc receptor Fc gamma RIIa on the platelet plasma membrane. Blood 2003; 102: 3637-45.
20 Rathore V, Stapleton MA, Hillery CA. et al. PECAM-1 negatively regulates GPIb/V/IX signaling in murine platelets. Blood 2003; 102: 3658-64.
21 Bodin S, Tronchere H, Payrastre B. Lipid rafts are critical membrane domains in blood platelet activation process. Biochim Biophys Acta 2003; 1610: 247-57.
22 Locke D, Chen H, Liu Y. et al. Lipid rafts orchestrate signaling by the platelet receptor glycoprotein VI. J Biol Chem 2001; 277: 18801-9.
23 Bodin S, Viala C, Ragab A. et al. A critical role of lipid rafts in the organization of a key Fc gammaRIIamediated signaling pathway in human platelets. Thromb Haemost 2003; 89: 318-30.
24 Sun QH, DeLisser HM, Zukowski MM. et al. Individually distinct Ig homology domains in PECAM-1 regulate homophilic binding and modulate receptor affinity. J Biol Chem 1996; 271: 11090-8.
25 Wong XM, Harbour SN, Wee JL. et al. Proteolytic cleavage of platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31) is regulated by a calmodulin-binding motif. FEBS Lett 2004; 568: 70-8.
26 Looney RJ, Abraham GN, Anderson CL. Human monocytes and U937 bear two distinct Fc receptors for IgG. J Immunol 1986; 136: 1641-7.
27 Rosenfeld SI, Ryan DH, Looney RJ. et al. Human Fc gamma receptors: stable inter-donor variation in quantitative expression on platelets correlates with functional responses. J Immunol 1987; 138: 2869-73.
28 Newman PJ, Hillery CA, Albrecht R. et al. Activation-dependent changes in human platelet PECAM-1: phosphorylation, cytoskeletal association, and surface membrane redistribution. J Cell Biol 1992; 119: 239-46.
29 Parton R, Hancock J. Caveolin and Ras function. Methods Enzymol 2001; 333: 172-83.
30 Del Conde I, Shrimpton CN, Thiagarajan P. et al. Tissue-factor bearing microvesicles arise from lipid rafts and fuse with activated platelets to initiate coagulation. Blood 2005; 106: 1604-11.
31 Gratzinger D, Canosa S, Engelhardt B. et al. Platelet endothelial cell adhesion molecule-1 modulates endothelial cell motility through the small G-protein Rho. FASEBJ 2003; 17: 1458-69.
32 Dorahy DJ, Lincz LF, Meldrun CJ. et al. Biochemical isolation of a membrane microdomain from resting platelets highly enriched in the plasma membrane glycoprotein CD36. Biochem J 1996; 319: 67-72.
33 Dorahy DJ, Burns GF. Active Lyn protein tyrosine kinase is selectively enriched within membrane microdomains of resting platelets. Biochem J 1998; 333: 373-9.
34 Wee JL, Jackson DE. The Ig-ITIM superfamily member PECAM-1 regulates the ‘outside-in’signaling properties of integrin αIIbβ3 in platelets. Blood 2005; 106: 3816-23.
35 Gousset K, Wolkers WF, Tsvetkova NM. et al. Evidence for a physiological role for membrane rafts in human platelets. J Cell Physiol 2002; 190: 117-28.
36 Heerklotz H. Triton promotes domain formation in lipid raft mixtures. BiophysJ 2002; 83: 2693-701.
37 Kwik J, Boyle S, Fooksman D. et al. Membrane cholesterol, lateral mobility, and the phosphatidylinositol 4,5-bisphosphate-dependent organization of cell actin. Proc Natl Acad Sci USA 2003; 100: 13964-9.
38 Wonerow P, Obergfell A, Wilde JI. et al. Differential role of glycolipid-enriched membrane microdomains in glycoprotein VI-and integrin-mediated phospholipase Cγ2 regulation in platelets. Biochem J 2002; 364: 755-65.
39 Hueber AO, Bernard AM, Herincs Z. et al. An essential role for membrane rafts in the initiation of Fas/ CD95-triggered cell death in mouse thymocytes. EMBO Rep 2002; 03: 190-6.
40 Hueber AO. Role of membrane microdomain rafts in TNFR-mediated signal transduction. Cell Death Differ 2003; 10: 7-9.
41 Delmas D, Rebe C, Micheau O. et al. Redistribution of CD95, DR4 and DR5 in rafts accounts for the synergistic toxicity of resveratrol and death receptor ligands in colon carcinoma cells. Oncogene 2004; 23: 8979-86.
42 Muppidi JR, Tschopp J, Siegel RM. Life and death decisions: secondary complexes and lipid rafts in TNF receptor family signal transduction. Immunity 2004; 21: 461-5.
43 Furne C, Corset V, Herincs Z. et al. The dependence receptor DCC requires lipid raft localization for cell death signaling. Proc Natl Acad Sci USA 2006; 103: 4128-33.