Role of Thrombospondin-1-Derived Peptide, 4N1K, in FGF-2-Induced Angiogenesis

Kanda, Shigeru; Shono, Takefumi; Tomasini-Johansson, Bianca R.; Klint, Peter; Saito, Yutaka

doi:10.1006/excr.1999.4622

Cited by 64 publications

(38 citation statements)

References 35 publications

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“…TSP1 strongly inhibits endothelial cell proliferation, as does CD148 (33,34). It also suppresses endothelial growth factor signaling, including VEGFR2 (40) and FGFR (41), but the mechanism of this suppression is incompletely understood. Further, the phenotype of TSP1 knockout mice suggests a role of TSP1 in negative regulation of epithelial cell proliferation (38).…”

Section: Discussionmentioning

confidence: 99%

Thrombospondin-1 acts as a ligand for CD148 tyrosine phosphatase

Takahashi¹,

Mernaugh²,

Friedman³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

109

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CD148 is a receptor-type protein tyrosine phosphatase that is expressed in several cell types, including vascular endothelial cells and duct epithelial cells. Growing evidence demonstrates a prominent role for CD148 in negative regulation of growth factor signals, suppressing cell proliferation and transformation. However, its extracellular ligand(s) remain unknown. To identify the ligand(s) of CD148, we introduced HA-tagged CD148 into cultured endothelial cells and then isolated its interacting extracellular protein(s) by biotin surface labeling and subsequent affinity purifications. The binding proteins were identified by mass spectrometry. Here we report that soluble thrombospondin-1 (TSP1) binds to the extracellular part of CD148 with high affinity and specificity, and its binding increases CD148 catalytic activity, leading to dephosphorylation of the substrate proteins. Consistent with these findings, introduction of CD148 conferred TSP1-mediated inhibition of cell growth to cells which lack CD148 and TSP1 inhibition of growth. Further, we demonstrate that TSP1-mediated inhibition of endothelial cell growth is antagonized by soluble CD148 ectodomain as well as by CD148 gene silencing. These findings provide evidence that CD148 functions as a receptor for TSP1 and mediates its inhibition of cell growth.growth factor signaling | EGF receptor | VEGF receptor | ERK

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Section: Discussionmentioning

confidence: 99%

Thrombospondin-1 acts as a ligand for CD148 tyrosine phosphatase

Takahashi¹,

Mernaugh²,

Friedman³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

109

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“…Thus, the NGVQYRN sequence probably is also exposed in the native protein and solvent-accessible. Recently, contrary to the initial report that the sequence NGVQYRN defines a minimal sequence within the TSP1 procollagen module capable of inhibiting corneal neovascularization (7), two other groups have not been able to show inhibitory activity for the peptide in either the CAM or corneal assay (60,61). Recombinant TSP1 procollagen module made in Escherichia coli also failed to inhibit vascularization in the CAM assay (61).…”

Section: Table IIImentioning

confidence: 94%

Physical Characterization of the Procollagen Module of Human Thrombospondin 1 Expressed in Insect Cells

Misenheimer

Huwiler

Annis

et al. 2000

Journal of Biological Chemistry

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Thrombospondin 1 (TSP1) is a homotrimeric glycoprotein composed of 150-kDa subunits connected by disulfide bridges. The procollagen module of thrombospondin 1 has been implicated in antiangiogenic activity. Procollagen modules are found in a number of extracellular proteins and are identifiable by 10 cysteines with characteristic spacing. We expressed and studied the procollagen module (C) of human TSP1, both by itself and in the context of the adjoining oligomerization sequence (o) and N-terminal module (N). The coding sequences were introduced into baculoviruses along with an N-terminal signal sequence and C-terminal polyhistidine tag. Proteins were purified from conditioned medium of infected insect cells by nickel-chelate chromatography. NoC is a disulfide bonded trimer and cleaves readily at a site of preferential proteolysis to yield monomeric N and trimeric oC. These are known properties of full-length TSP1. Mass spectroscopy indicated that C is N-glycosylated, and all 10 cysteine residues of C are in disulfides. By equilibrium ultracentrifugation, C is a monomer in physiological salt solution. Circular dichroism, intrinsic fluorescence, and differential scanning calorimetry experiments suggest that the stability of C is determined by the disulfides. The two tryptophans of C are in a polar, exposed environment as assessed by iodide fluorescence quenching and solvent perturbation. The oC far UV circular dichroism spectrum could be modeled as the sum of C and a coiled-coil oligomerization domain. The results indicate that the recombinant C folds autonomously into its native structure, and trimerization of the modules in TSP1 does not perturb their structures.

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“…The C-terminal G domain interacts with CD47. Some CD47-binding peptides derived from this domain inhibit angiogenesis [138] and CD47 is necessary for anti-angiogenic activities of TSP1 in the context of nitric oxide signaling [86]. Regulation of vascular smooth muscle cell contraction by TSP1.…”

Section: Future Directionsmentioning

confidence: 99%

Thrombospondins: from structure to therapeutics

Isenberg

Frazier

Roberts

2008

Cell. Mol. Life Sci.

118

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Thrombospondin-1 is secreted protein that modulates vascular cell behavior via several cell surface receptors. In vitro, nanomolar concentrations of thrombospondin-1 are required to alter endothelial and vascular smooth muscle cell adhesion, proliferation, motility, and survival. Yet, much lower levels of thrombospondin-1 are clearly functional in vivo. This discrepancy was explained with the discovery that the potency of thrombospondin-1 increases more than 100-fold in the presence of physiological levels of NO. Thrombospondin-1 binding to CD47 inhibits NO signaling by preventing cGMP synthesis and activation of its target cGMP-dependent protein kinase. This potent antagonism of NO signaling allows thrombospondin-1 to acutely constrict blood vessels, accelerate platelet aggregation and, if sustained, inhibit angiogenic responses. Acute antagonism of NO signaling by thrombospondin-1 is important for hemostasis but becomes detrimental for tissue survival of ischemic injuries. New therapeutic approaches targeting thrombospondin-1 or CD47 can improve recovery from ischemic injuries and overcome a deficit in NO-responsiveness in aging.

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Role of Thrombospondin-1-Derived Peptide, 4N1K, in FGF-2-Induced Angiogenesis

Cited by 64 publications

References 35 publications

Thrombospondin-1 acts as a ligand for CD148 tyrosine phosphatase

Thrombospondin-1 acts as a ligand for CD148 tyrosine phosphatase

Physical Characterization of the Procollagen Module of Human Thrombospondin 1 Expressed in Insect Cells

Thrombospondins: from structure to therapeutics

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