Molecular basis of antiangiogenic thrombospondin-1 type 1 repeat domain interactions with CD36. Arterioscler Thromb Vasc Biol 2013 Jul;33(7):1655-62
Date
05/04/2013Pubmed ID
23640500Pubmed Central ID
PMC3737738DOI
10.1161/ATVBAHA.113.301523Scopus ID
2-s2.0-84879103346 (requires institutional sign-in at Scopus site) 36 CitationsAbstract
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.
APPROACH AND 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.
Author List
Klenotic PA, Page RC, Li W, Amick J, Misra S, Silverstein RLAuthor
Roy L. Silverstein MD Professor in the Medicine department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Amino Acid SequenceAngiogenesis Inhibitors
Animals
Aorta
CD36 Antigens
Cell Movement
Cells, Cultured
Computer Simulation
Endothelial Cells
Human Umbilical Vein Endothelial Cells
Humans
Mice
Mice, Inbred C57BL
Models, Molecular
Molecular Sequence Data
Mutagenesis, Site-Directed
Neovascularization, Physiologic
Protein Binding
Protein Interaction Domains and Motifs
Protein Interaction Mapping
Recombinant Proteins
Surface Properties
Thrombospondin 1
Tissue Culture Techniques