title: Characterization of the interaction between endostatin short peptide and VEGF receptor 3.
abstract: Corneal angiogenesis and lymphangiogenesis are induced by vascular endothelial growth factors (VEGFs) signaling through its receptors VEGFR-1, -2, and -3. Endostatin is a peptide antagonist of these receptors that causes inhibition of bFGF-induced corneal angiogenesis and lymphangiogenesis. Here we show that binding of VEGF-C and endostatin to recombinant VEGFR-3 is competitive. Alignments of the primary amino acid sequences of VEGF-C and the C-terminal endostatin peptide (mEP: LEQKAASCHNSYIVLCIENSFMTSFSK) identified two conserved cysteine residues separated by seven amino acids. Peptides of VEGF-C and mEP containing these conserved residues bound to VEGFR-3. However, substitution of alanine for either of the cysteines in the mEP peptide perturbed the secondary structure, and this mutated peptide was unable to bind to VEGFR-3. Analysis by surface plasmon resonance demonstrated that the binding of the mEP peptide for recombinant VEGFR-3 had a Ka of 1.41 x 10⁷ M⁻¹ s⁻¹, Kd of 0.6718 s⁻¹, and a KD of 4.78 x 10⁻⁸ M. Characterization of the mechanism of endostatin binding to VEGFR-3 may lead to the development of novel therapies for lymphangiogenesis-related disorders, such as transplant rejection, lymphedema, and cancer metastasis.
authors.affiliation: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, University of Illinois at Chicago, Chicago, IL, USA.
authors.forename: Kyu-Yeon
authors.initials: KY
authors.lastname: Han
authors.forename: Dimitri T
authors.initials: DT
authors.lastname: Azar
authors.forename: Abdellah
authors.initials: A
authors.lastname: Sabri
authors.forename: Hyun
authors.initials: H
authors.lastname: Lee
authors.forename: Sandeep
authors.initials: S
authors.lastname: Jain
authors.forename: Bao-Shiang
authors.initials: BS
authors.lastname: Lee
authors.forename: Jin-Hong
authors.initials: JH
authors.lastname: Chang
mesh_terms: D000595:Amino Acid Sequence; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D017124:Conserved Sequence; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D043169:Endostatins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D016415:Sequence Alignment; D042582:Vascular Endothelial Growth Factor C / Q000737:chemistry / Q000378:metabolism*; D040321:Vascular Endothelial Growth Factor Receptor-3 / Q000378:metabolism*
chemical_list: D043169:Endostatins; D010455:Peptides; D042582:Vascular Endothelial Growth Factor C; D040321:Vascular Endothelial Growth Factor Receptor-3; D003545:Cysteine