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CD36 mediates the In vitro inhibitory effects of thrombospondin-1 on endothelial cells.

Dawson DW, Pearce SF, Zhong R, Silverstein RL, Frazier WA, Bouck NP - J. Cell Biol. (1997)

Bottom Line: Here we use both native TSP-1 and small antiangiogenic peptides derived from it to show that this inhibition is mediated by CD36, a transmembrane glycoprotein found on microvascular endothelial cells.Additional molecules known to bind CD36, including the IgM anti-CD36 antibody SM, oxidized (but not unoxidized) low density lipoprotein, and human collagen 1, mimicked TSP-1 by inhibiting the migration of human microvascular endothelial cells.Transfection of CD36-deficient human umbilical vein endothelial cells with a CD36 expression plasmid caused them to become sensitive to TSP-1 inhibition of their migration and tube formation.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology-Immunology and Robert H. Lurie Cancer Center, Northwestern University Medical School, Chicago, Illinois 60611, USA.

ABSTRACT
Thrombospondin-1 (TSP-1) is a naturally occurring inhibitor of angiogenesis that is able to make normal endothelial cells unresponsive to a wide variety of inducers. Here we use both native TSP-1 and small antiangiogenic peptides derived from it to show that this inhibition is mediated by CD36, a transmembrane glycoprotein found on microvascular endothelial cells. Both IgG antibodies against CD36 and glutathione-S-transferase-CD36 fusion proteins that contain the TSP-1 binding site blocked the ability of intact TSP-1 and its active peptides to inhibit the migration of cultured microvascular endothelial cells. In addition, antiangiogenic TSP-1 peptides inhibited the binding of native TSP-1 to solid phase CD36 and its fusion proteins, as well as to CD36-expressing cells. Additional molecules known to bind CD36, including the IgM anti-CD36 antibody SM, oxidized (but not unoxidized) low density lipoprotein, and human collagen 1, mimicked TSP-1 by inhibiting the migration of human microvascular endothelial cells. Transfection of CD36-deficient human umbilical vein endothelial cells with a CD36 expression plasmid caused them to become sensitive to TSP-1 inhibition of their migration and tube formation. This work demonstrates that endothelial CD36, previously thought to be involved only in adhesion and scavenging activities, may be essential for the inhibition of angiogenesis by thrombospondin-1.

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Related in: MedlinePlus

The relationship of small TSP-1 peptides to the 180-kD  monomer of TSP-1 and of GST–CD36 fusion proteins to the  whole CD36 receptor protein. Numbers indicate amino acid residues present in the peptide or fusion protein. Shading on the  whole CD36 molecule defines the minimal region of CD36 required for binding to TSP-1 (Pearce et al., 1995). Actual peptide  sequences and detailed description of the fusion proteins are included in Materials and Methods.
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Figure 1: The relationship of small TSP-1 peptides to the 180-kD monomer of TSP-1 and of GST–CD36 fusion proteins to the whole CD36 receptor protein. Numbers indicate amino acid residues present in the peptide or fusion protein. Shading on the whole CD36 molecule defines the minimal region of CD36 required for binding to TSP-1 (Pearce et al., 1995). Actual peptide sequences and detailed description of the fusion proteins are included in Materials and Methods.

Mentions: Purified human platelet TSP-1 was kindly provided by Jack Lawler (Harvard University, Cambridge, MA). TSP-1 peptides were synthesized, purified, and dialyzed as previously described (Tolsma et al., 1993) and included Col overlap, NGVQYRN representing TSP-1 amino acid residues 303–309; Mal III, SPWDIASVTAGGGVQKRSK representing TSP-1 amino acid residues 481–499 with the cysteines present in the native molecule replaced with alanines; and Mal III variant, SPWDIASTSAGGGVQRSK, containing Mal III residues with an altered VTCG sequence. The position of these peptides in the intact TSP-1 molecule is shown in Fig. 1. Angiostatin was kindly provided by Michael O'Reilly and Judah Folkman (Harvard University). Recombinant human basic fibroblast growth factor (bFGF) was purchased from R & D Systems Inc. (Minneapolis, MN), human type I collagen was from Collaborative Biomedical Products (Bedford, MA), and human LDL was from Sigma Chemical Co. LDL was oxidized by dialyzing 200 μg/ml LDL against 5 μM CuSO4 in PBS for 24 h at 37°C (Nicholson et al., 1995).


CD36 mediates the In vitro inhibitory effects of thrombospondin-1 on endothelial cells.

Dawson DW, Pearce SF, Zhong R, Silverstein RL, Frazier WA, Bouck NP - J. Cell Biol. (1997)

The relationship of small TSP-1 peptides to the 180-kD  monomer of TSP-1 and of GST–CD36 fusion proteins to the  whole CD36 receptor protein. Numbers indicate amino acid residues present in the peptide or fusion protein. Shading on the  whole CD36 molecule defines the minimal region of CD36 required for binding to TSP-1 (Pearce et al., 1995). Actual peptide  sequences and detailed description of the fusion proteins are included in Materials and Methods.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2141641&req=5

Figure 1: The relationship of small TSP-1 peptides to the 180-kD monomer of TSP-1 and of GST–CD36 fusion proteins to the whole CD36 receptor protein. Numbers indicate amino acid residues present in the peptide or fusion protein. Shading on the whole CD36 molecule defines the minimal region of CD36 required for binding to TSP-1 (Pearce et al., 1995). Actual peptide sequences and detailed description of the fusion proteins are included in Materials and Methods.
Mentions: Purified human platelet TSP-1 was kindly provided by Jack Lawler (Harvard University, Cambridge, MA). TSP-1 peptides were synthesized, purified, and dialyzed as previously described (Tolsma et al., 1993) and included Col overlap, NGVQYRN representing TSP-1 amino acid residues 303–309; Mal III, SPWDIASVTAGGGVQKRSK representing TSP-1 amino acid residues 481–499 with the cysteines present in the native molecule replaced with alanines; and Mal III variant, SPWDIASTSAGGGVQRSK, containing Mal III residues with an altered VTCG sequence. The position of these peptides in the intact TSP-1 molecule is shown in Fig. 1. Angiostatin was kindly provided by Michael O'Reilly and Judah Folkman (Harvard University). Recombinant human basic fibroblast growth factor (bFGF) was purchased from R & D Systems Inc. (Minneapolis, MN), human type I collagen was from Collaborative Biomedical Products (Bedford, MA), and human LDL was from Sigma Chemical Co. LDL was oxidized by dialyzing 200 μg/ml LDL against 5 μM CuSO4 in PBS for 24 h at 37°C (Nicholson et al., 1995).

Bottom Line: Here we use both native TSP-1 and small antiangiogenic peptides derived from it to show that this inhibition is mediated by CD36, a transmembrane glycoprotein found on microvascular endothelial cells.Additional molecules known to bind CD36, including the IgM anti-CD36 antibody SM, oxidized (but not unoxidized) low density lipoprotein, and human collagen 1, mimicked TSP-1 by inhibiting the migration of human microvascular endothelial cells.Transfection of CD36-deficient human umbilical vein endothelial cells with a CD36 expression plasmid caused them to become sensitive to TSP-1 inhibition of their migration and tube formation.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology-Immunology and Robert H. Lurie Cancer Center, Northwestern University Medical School, Chicago, Illinois 60611, USA.

ABSTRACT
Thrombospondin-1 (TSP-1) is a naturally occurring inhibitor of angiogenesis that is able to make normal endothelial cells unresponsive to a wide variety of inducers. Here we use both native TSP-1 and small antiangiogenic peptides derived from it to show that this inhibition is mediated by CD36, a transmembrane glycoprotein found on microvascular endothelial cells. Both IgG antibodies against CD36 and glutathione-S-transferase-CD36 fusion proteins that contain the TSP-1 binding site blocked the ability of intact TSP-1 and its active peptides to inhibit the migration of cultured microvascular endothelial cells. In addition, antiangiogenic TSP-1 peptides inhibited the binding of native TSP-1 to solid phase CD36 and its fusion proteins, as well as to CD36-expressing cells. Additional molecules known to bind CD36, including the IgM anti-CD36 antibody SM, oxidized (but not unoxidized) low density lipoprotein, and human collagen 1, mimicked TSP-1 by inhibiting the migration of human microvascular endothelial cells. Transfection of CD36-deficient human umbilical vein endothelial cells with a CD36 expression plasmid caused them to become sensitive to TSP-1 inhibition of their migration and tube formation. This work demonstrates that endothelial CD36, previously thought to be involved only in adhesion and scavenging activities, may be essential for the inhibition of angiogenesis by thrombospondin-1.

Show MeSH
Related in: MedlinePlus