Biological Activity and Phosphorylation Sites of the Bacterially Expressed Cytosolic Domain of the KDR VEGF-Receptor

Dougher-Vermazen, Maureen; Hulmes, Jeffrey D.; Bőhlen, Peter; Terman, Bruce I.

doi:10.1006/bbrc.1994.2726

Cited by 109 publications

(67 citation statements)

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“…In yeast two hybrid system, we show that the interaction between Grb10 and KDR is dependent on the receptor tyrosine kinase activity and on the integrity of the SH2 domain of Grb10. Grb10 SH2 domain recognizes speci®c tyrosine phosphorylated sequences in several tyrosine kinase receptors (Frantz et al, 1997;Stein et al, 1996;Wang et al, 1999 (Dougher-Vermazen et al, 1994;Wu et al, 2000). However, mutation of several KDR tyrosine residues, including autophosphorylation sites, did not allow us to identify the binding site of Grb10 on KDR (data not shown).…”

Section: Discussionmentioning

confidence: 96%

The adapter protein, Grb10, is a positive regulator of vascular endothelial growth factor signaling

Giorgetti‐Peraldi

Murdaca

et al. 2001

Oncogene

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Section: Discussionmentioning

confidence: 96%

The adapter protein, Grb10, is a positive regulator of vascular endothelial growth factor signaling

Giorgetti‐Peraldi

Murdaca

et al. 2001

Oncogene

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“…Dougher- Vermazen et al (15) reported that in a bacteria expression system, Y951, Y996, Y1054, and Y1059 on KDR were phosphorylated. On the other hand, Cunningham et al (16) reported that in the yeast system, Y801 and Y1175 on KDR were phosphorylated.…”

Section: Discussionmentioning

confidence: 99%

Essential role of Flk-1 (VEGF receptor 2) tyrosine residue 1173 in vasculogenesis in mice

Sakurai

Ohgimoto²,

Kataoka³

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

286

250

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Flk-1 (human counterpart, KDR) tyrosine kinase, which is one of the two VEGF receptors, is crucial for vascular development. Recently, we showed that, among tyrosine residues of KDR, tyrosine residues 1175 (Y1175, corresponding to Y1173 in murine Flk-1) and Y1214 (Y1212 in Flk-1) are autophosphorylated in response to VEGF, and that Y1175 is important for VEGF-dependent phospholipase C␥͞PKC͞mitogen-activated protein kinase activation leading to DNA synthesis in cultured endothelial cells. However, the importance of these tyrosine residues in Flk-1͞KDR in vivo is not yet known. To examine the role of these Flk-1 tyrosine residues in vivo, we generated knock-in mice substituting Y1173 and Y1212 of the Flk-1 gene with phenylalanine, respectively. As a result, Flk-1 1173F homozygous mice died between embryonic days 8.5 and 9.5 without any organized blood vessels or yolk sac blood islands, and hematopoietic progenitors were severely reduced, similar to the case of Flk-1 null mice. In contrast, Flk-1 1212F homozygous mice were viable and fertile. These results suggest that the signaling via Y1173 of Flk-1 is essential for endothelial and hematopoietic development during embryogenesis.Flk-1͞KDR ͉ single tyrosine-phenylalanine mutation ͉ tyrosine kinase receptor V EGF is essential for many angiogenic processes both in normal and pathological conditions (1, 2). VEGF binds two tyrosine kinase receptors, Flt-1 (also known as VEGF receptor 1) (3, 4) and Flk-1 (also known as VEGF receptor 2, KDR as human counterpart) (5-7) with high affinity. Flt-1-and Flk-1-deficient mice both die in utero between embryonic days (E) 8.5 and E9.5 but have different phenotypes. Flt-1-deficient embryos showed an overgrowth of endothelial cells and disorganization of blood vessels (8). Moreover, Flt-1 tyrosine kinase-deficient mice showed normal vascular development (9), suggesting that the Flt-1 extracellular domain acts as a negative regulator of VEGF, and that Flt-1 tyrosine kinase is not necessary for vasculogenesis during development. On the other hand, Flk-1-deficient mice lack both mature endothelial and hematopoietic cells, indicating that Flk-1 is crucial for vascular development (10).Ligand binding to Flk-1͞KDR induces autophosphorylation of Flk-1͞KDR intracellular tyrosine residues and activates several signaling pathways, leading to cell proliferation, survival, migration, and permeability (11). Recently, we have shown that tyrosine residues 1175 (Y1175) and Y1214, but not Y801, on KDR are highly autophosphorylated in response to VEGF, and that Y1175 is crucial for VEGF-dependent cell proliferation via the phospholipase C␥ (PLC␥)͞PKC͞mitogen-activated protein kinase (MAPK) pathway in cultured endothelial cells (12). However, the importance of these tyrosine residues in Flk-1͞KDR in vivo remains to be elucidated.To develop a pharmaceutical drug(s) that efficiently suppresses angiogenesis in diseases such as cancer, it is important to identify the tyrosine residue(s) that is involved in the critical signaling pathway via Flk-1͞KDR for ...

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“…Major autophosphorylation sites of VEGFR-2 are located in the kinase insert domain (Tyr951/996) and in the tyrosine kinase catalytic domain (Tyr1054/1059) (Dougher-Vermazen et al, 1994). In endothelial cells overexpressing VEGFR-2, activation of the receptor leads to a rapid recruitment of the adaptor proteins Shc, Grb2 and Nck, and protein tyrosine phosphatases SHP-1 and SHP-2 (Kroll and Waltenberger, 1997).…”

Section: Vegfr-2mentioning

confidence: 99%

Vascular endothelial growth factor receptors in the regulation of angiogenesis and lymphangiogenesis

2000

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Biological Activity and Phosphorylation Sites of the Bacterially Expressed Cytosolic Domain of the KDR VEGF-Receptor

Cited by 109 publications

References 0 publications

The adapter protein, Grb10, is a positive regulator of vascular endothelial growth factor signaling

The adapter protein, Grb10, is a positive regulator of vascular endothelial growth factor signaling

Essential role of Flk-1 (VEGF receptor 2) tyrosine residue 1173 in vasculogenesis in mice

Vascular endothelial growth factor receptors in the regulation of angiogenesis and lymphangiogenesis

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