title: Biosynthesis of vascular endothelial growth factor-D involves proteolytic processing which generates non-covalent homodimers.
abstract: Vascular endothelial growth factor-D (VEGF-D) binds and activates the endothelial cell tyrosine kinase receptors VEGF receptor-2 (VEGFR-2) and VEGF receptor-3 (VEGFR-3), is mitogenic for endothelial cells, and shares structural homology and receptor specificity with VEGF-C. The primary translation product of VEGF-D has long N- and C-terminal polypeptide extensions in addition to a central VEGF homology domain (VHD). The VHD of VEGF-D is sufficient to bind and activate VEGFR-2 and VEGFR-3. Here we report that VEGF-D is proteolytically processed to release the VHD. Studies in 293EBNA cells demonstrated that VEGF-D undergoes N- and C-terminal cleavage events to produce numerous secreted polypeptides including a fully processed form of M(r) approximately 21,000 consisting only of the VHD, which is predominantly a non-covalent dimer. Biosensor analysis demonstrated that the VHD has approximately 290- and approximately 40-fold greater affinity for VEGFR-2 and VEGFR-3, respectively, compared with unprocessed VEGF-D. In situ hybridization demonstrated that embryonic lung is a major site of expression of the VEGF-D gene. Processed forms of VEGF-D were detected in embryonic lung indicating that VEGF-D is proteolytically processed in vivo.
authors.affiliation: Ludwig Institute for Cancer Research, Royal Melbourne Hospital, Parkville, Victoria 3050, Australia. steven.stacker@ludwig.edu.au
authors.forename: S A
authors.initials: SA
authors.lastname: Stacker
authors.forename: K
authors.initials: K
authors.lastname: Stenvers
authors.forename: C
authors.initials: C
authors.lastname: Caesar
authors.forename: A
authors.initials: A
authors.lastname: Vitali
authors.forename: T
authors.initials: T
authors.lastname: Domagala
authors.forename: E
authors.initials: E
authors.lastname: Nice
authors.forename: S
authors.initials: S
authors.lastname: Roufail
authors.forename: R J
authors.initials: RJ
authors.lastname: Simpson
authors.forename: R
authors.initials: R
authors.lastname: Moritz
authors.forename: T
authors.initials: T
authors.lastname: Karpanen
authors.forename: K
authors.initials: K
authors.lastname: Alitalo
authors.forename: M G
authors.initials: MG
authors.lastname: Achen
mesh_terms: D002455:Cell Division; D002460:Cell Line; D019281:Dimerization; D016228:Endothelial Growth Factors / Q000096:biosynthesis*; D004730:Endothelium, Vascular / Q000378:metabolism; D006801:Humans; D017403:In Situ Hybridization; D007202:Indicators and Reagents; D009842:Oligopeptides; D010455:Peptides; D011499:Protein Processing, Post-Translational; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism; D017978:Receptors, Growth Factor / Q000378:metabolism; D011975:Receptors, Mitogen / Q000378:metabolism; D040262:Receptors, Vascular Endothelial Growth Factor; D042643:Vascular Endothelial Growth Factor D; D040321:Vascular Endothelial Growth Factor Receptor-3
chemical_list: D016228:Endothelial Growth Factors; D007202:Indicators and Reagents; D009842:Oligopeptides; D010455:Peptides; D011956:Receptors, Cell Surface; D017978:Receptors, Growth Factor; D011975:Receptors, Mitogen; D042643:Vascular Endothelial Growth Factor D; C085706:FLAG peptide; D020794:Receptor Protein-Tyrosine Kinases; D040262:Receptors, Vascular Endothelial Growth Factor; D040321:Vascular Endothelial Growth Factor Receptor-3