title: The propeptides of VEGF-D determine heparin binding, receptor heterodimerization, and effects on tumor biology.
abstract: VEGF-D is an angiogenic and lymphangiogenic glycoprotein that can be proteolytically processed generating various forms differing in subunit composition due to the presence or absence of N- and C-terminal propeptides. These propeptides flank the central VEGF homology domain, that contains the binding sites for VEGF receptors (VEGFRs), but their biological functions were unclear. Characterization of propeptide function will be important to clarify which forms of VEGF-D are biologically active and therefore clinically relevant. Here we use VEGF-D mutants deficient in either propeptide, and in the capacity to process the remaining propeptide, to monitor the functions of these domains. We report for the first time that VEGF-D binds heparin, and that the C-terminal propeptide significantly enhances this interaction (removal of this propeptide from full-length VEGF-D completely prevents heparin binding). We also show that removal of either the N- or C-terminal propeptide is required for VEGF-D to drive formation of VEGFR-2/VEGFR-3 heterodimers which have recently been shown to positively regulate angiogenic sprouting. The mature form of VEGF-D, lacking both propeptides, can also promote formation of these receptor heterodimers. In a mouse tumor model, removal of only the C-terminal propeptide from full-length VEGF-D was sufficient to enhance angiogenesis and tumor growth. In contrast, removal of both propeptides is required for high rates of lymph node metastasis. The findings reported here show that the propeptides profoundly influence molecular interactions of VEGF-D with VEGF receptors, co-receptors, and heparin, and its effects on tumor biology.
authors: Tumour Angiogenesis Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria 3002, Australia; Ludwig Institute for Cancer Research, Royal Melbourne Hospital, Parkville, Victoria 3050, Australia.
authors: Nicole C
authors: NC
authors: Harris
authors: Tumour Angiogenesis Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria 3002, Australia.
authors: Natalia
authors: N
authors: Davydova
authors: Tumour Angiogenesis Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria 3002, Australia.
authors: Sally
authors: S
authors: Roufail
authors: Tumour Angiogenesis Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria 3002, Australia.
authors: Sophie
authors: S
authors: Paquet-Fifield
authors: Ludwig Institute for Cancer Research, Royal Melbourne Hospital, Parkville, Victoria 3050, Australia.
authors: Karri
authors: K
authors: Paavonen
authors: Tumour Angiogenesis Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria 3002, Australia.
authors: Tara
authors: T
authors: Karnezis
authors: Tumour Angiogenesis Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria 3002, Australia.
authors: You-Fang
authors: YF
authors: Zhang
authors: Tumour Angiogenesis Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria 3002, Australia.
authors: Teruhiko
authors: T
authors: Sato
authors: Ludwig Institute for Cancer Research, Royal Melbourne Hospital, Parkville, Victoria 3050, Australia.
authors: Julie
authors: J
authors: Rothacker
authors: Ludwig Institute for Cancer Research, Royal Melbourne Hospital, Parkville, Victoria 3050, Australia.
authors: Edouard C
authors: EC
authors: Nice
authors: Tumour Angiogenesis Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria 3002, Australia; Ludwig Institute for Cancer Research, Royal Melbourne Hospital, Parkville, Victoria 3050, Australia; Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria 3050, Australia.
authors: Steven A
authors: SA
authors: Stacker
authors: Tumour Angiogenesis Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria 3002, Australia; Ludwig Institute for Cancer Research, Royal Melbourne Hospital, Parkville, Victoria 3050, Australia; Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria 3050, Australia. Electronic address: Marc.achen@petermac.org.
authors: Marc G
authors: MG
authors: Achen
mesh_terms: D000818:Animals; D002460:Cell Line; D002846:Chromatography, Affinity; D042783:Endothelial Cells / Q000378:metabolism; D005260:Female; D006493:Heparin / Q000737:chemistry*; D006801:Humans; D042583:Lymphangiogenesis; D008207:Lymphatic Metastasis; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009368:Neoplasm Transplantation; D009374:Neoplasms, Experimental / Q000098:blood supply / Q000473:pathology; D009389:Neovascularization, Pathologic / Q000378:metabolism; D039921:Neuropilins / Q000378:metabolism; D011485:Protein Binding; D055503:Protein Multimerization; D011498:Protein Precursors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D017434:Protein Structure, Tertiary; D017384:Sequence Deletion; D042643:Vascular Endothelial Growth Factor D / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000737:chemistry / Q000378:metabolism*; D040321:Vascular Endothelial Growth Factor Receptor-3 / Q000737:chemistry / Q000378:metabolism*
chemical_list: D039921:Neuropilins; D011498:Protein Precursors; D042643:Vascular Endothelial Growth Factor D; D006493:Heparin; D040301:Vascular Endothelial Growth Factor Receptor-2; D040321:Vascular Endothelial Growth Factor Receptor-3
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