The carboxyl terminus controls ligand-dependent activation of VEGFR-2 and its signaling.

Vascular endothelial growth factor receptor-2 (VEGFR-2/FLK-1) is a receptor tyrosine kinase whose activation stimulates angiogenesis. We recently generated a chimeric VEGFR-2 in which the extracellular domain of VEGFR-2 was replaced with the extracellular domain of human colony stimulating factor-1 receptor and expressed in endothelial cells. To study the contribution of the carboxyl terminus to activation of VEGFR-2, we created a panel of truncated receptors in which the carboxyl terminus of VEGFR-2 was progressively deleted. Removal of the entire carboxyl terminus eliminated activation of VEGFR-2, its ability to activate signaling proteins, and its ability to stimulate cell proliferation. The carboxyl terminus-deleted VEGFR-2 exhibited impaired ligand-dependent down-regulation and inhibited the activation of wild-type receptor in a dominant-negative fashion. Furthermore, introducing the carboxyl terminus of another receptor, i.e., VEGFR-1, restored the ligand-dependent activation of the carboxyl terminus-deleted VEGFR-2 and its ability to stimulate cell proliferation. Our findings suggest that the carboxyl terminus of VEGFR-2 plays a critical role in VEGFR-2 activation, its ability to activate signaling proteins, and its ability to induce biological responses. The presence of at least 57 amino acids at the carboxyl terminus of VEGFR-2 are required for VEGFR-2 activation. Thus, we propose that the carboxyl terminus is required for activation of VEGFR-2, and absence of the carboxyl terminus renders VEGFR-2 inactive.