title: Molecular basis of antiangiogenic thrombospondin-1 type 1 repeat domain interactions with CD36.
abstract: OBJECTIVE
  Antiangiogenic activity of thrombospondin-1 and related proteins is mediated by interactions between thrombospondin type 1 repeat (TSR) domains and the CD36, LIMP-2, Emp sequence homology (CLESH) domain of the endothelial cell receptor CD36. We sought to characterize key molecular determinants of the interaction between thrombospondin-1 TSR domains and the CD36 CLESH domain.
  
  
  RESULTS
  Recombinant thrombospondin-1 TSR2 and TSR(2,3) constructs inhibited microvascular endothelial cell migration, microvascular endothelial cell tube formation, and vessel sprouting in aortic ring assays. Interaction with CD36 CLESH decoy peptides negated these effects. Mutational analyses identified a cluster of residues that confer positive charge to the TSR2 surface and mediate interaction with CD36 CLESH. Antiangiogenic activity was significantly reduced by charge-neutralizing mutations of the Arg-Trp ladder in TSR2, but not TSR3. Additionally, I438 and K464 of TSR2 were shown to be required for CD36 CLESH binding to TSR2. A complementary acidic cluster within CD36 CLESH is also required for antiangiogenic activity.
  
  
  CONCLUSIONS
  Thrombospondin-1 interacts with CD36 CLESH through electrostatic interactions mediated by a positively charged TSR2 surface and multiple negatively charged CD36 CLESH residues. Two key residues serve as specificity determinants that identify other TSR domains that interact with CD36 CLESH.
authors: Department of Molecular Cardiology, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA.
authors: Philip A
authors: PA
authors: Klenotic
authors: Richard C
authors: RC
authors: Page
authors: Wei
authors: W
authors: Li
authors: Joseph
authors: J
authors: Amick
authors: Saurav
authors: S
authors: Misra
authors: Roy L
authors: RL
authors: Silverstein
mesh_terms: D000595:Amino Acid Sequence; D020533:Angiogenesis Inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001011:Aorta / Q000378:metabolism*; D018955:CD36 Antigens / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002465:Cell Movement; D002478:Cells, Cultured; D003198:Computer Simulation; D042783:Endothelial Cells / Q000378:metabolism*; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D018919:Neovascularization, Physiologic; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D025941:Protein Interaction Mapping; D011994:Recombinant Proteins / Q000378:metabolism; D013499:Surface Properties; D019700:Thrombospondin 1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D046509:Tissue Culture Techniques
chemical_list: D020533:Angiogenesis Inhibitors; D018955:CD36 Antigens; D011994:Recombinant Proteins; D019700:Thrombospondin 1
keywords: CD36; angiogenesis; thrombospondin-1
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