TY - JOUR
T1 - A quantitative proteomic analysis of growth factor-induced compositional changes in lipid rafts of human smooth muscle cells
AU - MacLellan, Dawn L.
AU - Steen, Hanno
AU - Adam, Rosalyn M.
AU - Garlick, Monica
AU - Zurakowski, David
AU - Gygi, Steven P.
AU - Freeman, Michael R.
AU - Solomon, Keith R.
PY - 2005/12
Y1 - 2005/12
N2 - Signals that promote proliferation and migration of smooth muscle cells (SMC) have been implicated in pathologic growth of hollow organs. Members of the platelet-derived growth factor (PDGF) family, potent mitogens and motility factors for SMC, have been shown to signal through cholesterol-enriched lipid rafts. We recently demonstrated that PDGF-stimulated DNA synthesis in urinary tract SMC was dependent on the integrity of lipid rafts. Despite its known ability to rapidly alter discrete proteins within rafts, the effect of PDGF on overall raft protein composition is unknown. In this study, we employed isotope coded affinity tag (ICAT) analysis to evaluate PDGF-induced protein changes in lipid rafts of primary culture human SMC. Following acute (i.e., 15 min) exposure of SMC to PDGF, 23 proteins increased in rafts >20%. In contrast, raft localization of only three proteins increased after 12 h of PDGF treatment. Among the proteins that increased at 15 min were the glycophosphatidylinositol- anchored proteins Thy-1, 5′-nucleotidase, and CD55, the cytoskeletal proteins actin, actinin, tropomyosin-3 and -4, and the endocytosis-related proteins clathrin and β-adaptin. In addition, eight Rho family members were localized to rafts by ICAT analysis. Collectively, these observations suggest a role for lipid rafts in regulation of PDGF-stimulated changes in the cytoskeleton.
AB - Signals that promote proliferation and migration of smooth muscle cells (SMC) have been implicated in pathologic growth of hollow organs. Members of the platelet-derived growth factor (PDGF) family, potent mitogens and motility factors for SMC, have been shown to signal through cholesterol-enriched lipid rafts. We recently demonstrated that PDGF-stimulated DNA synthesis in urinary tract SMC was dependent on the integrity of lipid rafts. Despite its known ability to rapidly alter discrete proteins within rafts, the effect of PDGF on overall raft protein composition is unknown. In this study, we employed isotope coded affinity tag (ICAT) analysis to evaluate PDGF-induced protein changes in lipid rafts of primary culture human SMC. Following acute (i.e., 15 min) exposure of SMC to PDGF, 23 proteins increased in rafts >20%. In contrast, raft localization of only three proteins increased after 12 h of PDGF treatment. Among the proteins that increased at 15 min were the glycophosphatidylinositol- anchored proteins Thy-1, 5′-nucleotidase, and CD55, the cytoskeletal proteins actin, actinin, tropomyosin-3 and -4, and the endocytosis-related proteins clathrin and β-adaptin. In addition, eight Rho family members were localized to rafts by ICAT analysis. Collectively, these observations suggest a role for lipid rafts in regulation of PDGF-stimulated changes in the cytoskeleton.
UR - http://www.scopus.com/inward/record.url?scp=29544452310&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=29544452310&partnerID=8YFLogxK
U2 - 10.1002/pmic.200500044
DO - 10.1002/pmic.200500044
M3 - Article
C2 - 16267816
AN - SCOPUS:29544452310
SN - 1615-9853
VL - 5
SP - 4733
EP - 4742
JO - Proteomics
JF - Proteomics
IS - 18
ER -