A VEGF-A splice variant defective for heparan sulfate and neuropilin-1 binding shows attenuated signaling through VEGFR-2.
The development of functional blood and lymphatic vessels requires spatio-temporal coordination of the production and release of growth factors such as vascular endothelial growth factors (VEGFs). VEGF family proteins are produced in multiple isoforms with distinct biological properties and bind to three types of VEGF receptors. A VEGF-A splice variant, VEGF-A(165)b, has recently been isolated from kidney epithelial cells. This variant is identical to VEGF-A(165) except for the last six amino acids encoded by an alternative exon. VEGF-A(165)b and VEGF-A(165) bind VEGF receptors 1 and 2 with similar affinity. VEGF-A(165)b elicits drastically reduced activity in angiogenesis assays and even counteracts signaling by VEGF-A(165). VEGF-A(165)b weakly binds to heparan sulfate and does not interact with neuropilin-1, a coreceptor for VEGF receptor 2. To determine the molecular basis for altered signaling by VEGF-A(165)b we measured VEGF receptor 2 and ERK kinase activity in endothelial cells in culture. VEGF-A(165) induced strong and sustained activation of VEGF receptor 2 and ERK-1 and -2, while activation by VEGF-A(165)b was only weak and transient. Taken together these data show that VEGF-A(165)b has attenuated signaling potential through VEGF receptor 2 defining this new member of the VEGF family as a partial receptor agonist.
Cébe Suarez
S
S
Paul Scherrer Institut, Laboratory of Biomolecular Research, Molecular Cell Biology, 5232 Villigen-PSI, Switzerland.
Pieren
M
M
Cariolato
L
L
Arn
S
S
Hoffmann
U
U
Bogucki
A
A
Manlius
C
C
Wood
J
J
Ballmer-Hofer
K
K
D017398:Alternative Splicing; D000818:Animals; D002460:Cell Line; D002642:Chick Embryo; D049033:Chorioallantoic Membrane / Q000098:blood supply* / Q000378:metabolism; D006497:Heparan Sulfate / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D018919:Neovascularization, Physiologic; D039942:Neuropilin-1 / Q000378:metabolism*; D011485:Protein Binding; D020033:Protein Isoforms / Q000494:pharmacology; D015398:Signal Transduction; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics* / Q000493:pharmacokinetics / Q000494:pharmacology; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000378:metabolism; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism*
D006497:Heparan Sulfate; D039942:Neuropilin-1; D020033:Protein Isoforms; D042461:Vascular Endothelial Growth Factor A; D040281:Vascular Endothelial Growth Factor Receptor-1; D040301:Vascular Endothelial Growth Factor Receptor-2