Thrombospondin-1-induced vascular smooth muscle cell chemotaxis: the role of the type 3 repeat and carboxyl terminal domains.
Thrombospondin-1 (TSP-1), an acute phase reactant implicated in vascular disease, is a matricellular glycoprotein with six domains that confer different functions. The authors have shown TSP-1 induces vascular smooth muscle cell (VSMC) chemotaxis via extracellular signal-regulated kinases-1 and -2 (ERK) and p38 kinase (p38) and that a fusion protein of the carboxyl terminal (COOH) and type 3 repeat (T3) domains independently induce VSMC chemotaxis. The purpose of this study was to determine whether COOH-, T3-induced VSMC chemotaxis, or both, is dependent upon ERK or p38 activation. To determine if the T3, COOH, or type 2 repeat domain (T2, control domain not associated with chemotaxis) activate ERK, p38, or both, VSMCs were exposed to each fusion protein (20 microg/ml for 15, 30, 60, or 120 min), serum-free media (SFM, negative control), or TSP-1 (20 microg/ml for 30 min, positive control). Western immunoblotting was performed for activation studies. Using a microchemotaxis chamber, VSMCs pre-incubated in SFM, DMSO (vehicle control), PD98059 (10 microM), or SB202190 (10 microM) were exposed to each domain, TSP-1, or SFM. After 4 h (37 degrees C), migrated VSMCs were recorded as cells/five fields (400 x) and analyzed by paired t-test. ERK was activated by T2, T3, and COOH. However, p38 was activated by T3 and COOH, but not T2. T3 and COOH-induced VSMC chemotaxis were inhibited by PD98059 or SB202190, but more completely by SB202190. The T2 domain had no effect on VSMC chemotaxis. These results suggest activation of the p38 pathway may be more specific than ERK for COOH- and T3-induced VSMC chemotaxis.
Lee
Taeseung
T
Section of Vascular Surgery, Yale University School of Medicine, New Haven, Connecticut 06520, USA.
Nesselroth
Susan M
SM
Olson
Eric T
ET
Esemuede
Nowokere
N
Lawler
Jack
J
Sumpio
Bauer E
BE
Gahtan
Vivian
V
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002633:Chemotaxis / Q000502:physiology*; D004789:Enzyme Activation; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000201:enzymology / Q000502:physiology*; D019700:Thrombospondin 1 / Q000737:chemistry / Q000502:physiology*; D048051:p38 Mitogen-Activated Protein Kinases
D019700:Thrombospondin 1; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases