Identification of Vascular Endothelial Growth Factor Determinants for Binding KDR and FLT-1 Receptors

Keyt, Bruce A.; Nguyen, Hung Van; Berleau, Lea T.; Duarte, Carlos M.; Park, Jeanie; Chen, Helen; Ferrara, Napoleone

doi:10.1074/jbc.271.10.5638

Cited by 437 publications

(341 citation statements)

References 58 publications

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“…Ten ng/ml of VEGF was added to the cultures, which were further incubated for 15 min. After the incubation, cells were ®xed and analysed previous reports (Waltenberger et al, 1994;Keyt et al, 1996;Kroll et al, 1997;Bernatchez et al, 1999), and further con®rms that VEGF stimulates DNA synthesis of ECs preferentially via KDR. KDR-mediated signal preferentially modulates DNA synthesis of HUVECs via the activation of ERK1/2.…”

Section: Discussionmentioning

confidence: 93%

“…It has been reported that KDR but not Flt-1 plays a principal role in the transduction of VEGF-mediated e ects in porcine aortic ECs lacking endogenous VEGF receptors which are transfected with either Flt-1 or KDR gene (Waltenberger et al, 1994;Kroll et al, 1997). The importance of KDR/Flk-1 is further emphasized by using mutant VEGF which only binds to KDR (Keyt et al, 1996) or KDR antisense oligonucleotides (Bernatchez et al, 1999). However, Flt-1 does play a role in VEGF-stimulated migration of monocytes/ macrophages which express only Flt-1 (Shen et al, 1993;Barleon et al, 1996;Clauss et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

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Roles of two VEGF receptors, Flt-1 and KDR, in the signal transduction of VEGF effects in human vascular endothelial cells

Kanno¹,

Oda²,

Abé³

et al. 2000

Oncogene

260

175

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Section: Discussionmentioning

confidence: 93%

Section: Introductionmentioning

confidence: 99%

Roles of two VEGF receptors, Flt-1 and KDR, in the signal transduction of VEGF effects in human vascular endothelial cells

Kanno¹,

Oda²,

Abé³

et al. 2000

Oncogene

260

175

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“…Exon 1 plus 4 amino acids of exon 2 encodes the 26-residue signal peptide, cleaved during secretion. B, The final form of VEGF is a dimer, containing two of the above sequences covalently linked in an antiparallel orientation by cysteine-cysteine bonds; this makes VEGF bivalent, and VEGF receptors (VEGFR1, R2) bind in the regions encoded by exons 3 and 4 61 . Neuropilins (NRPs) and heparan sulfate (HS) chains bind VEGF in the region encoded by exons 6, 7 and 8a; thus isoforms have different receptor-binding characteristics depending on the encoding exons included.…”

Section: Vegf-a Isoforms Exhibit Differential Binding To Vegfrs and Nrpsmentioning

confidence: 99%

VEGF-A121a binding to Neuropilins – A concept revisited

Sarabipour

Gabhann

2017

Cell Adhesion & Migration

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All known splice isoforms of vascular endothelial growth factor A (VEGF-A) can bind to the receptor tyrosine kinases VEGFR-1 and VEGFR-2. We focus here on VEGF-A121a and VEGF-A165a, two of the most abundant VEGF-A splice isoforms in human tissue1, and their ability to bind the Neuropilin co-receptors NRP1 and NRP2. The Neuropilins are key vascular, immune, and nervous system receptors on endothelial cells, neuronal axons, and regulatory T cells respectively. They serve as co-receptors for the Plexins in Semaphorin binding on neuronal and vascular endothelial cells, and for the VEGFRs in VEGF binding on vascular and lymphatic endothelial cells, and thus regulate the initiation and coordination of cell signaling by Semaphorins and VEGFs.2 There is conflicting evidence in the literature as to whether only heparin-binding VEGF-A isoforms – that is, isoforms with domains encoded by exons 6 and/or 7 plus 8a – bind to Neuropilins on endothelial cells. While it is clear that VEGF-A165a binds to both NRP1 and NRP2, published studies do not all agree on the ability of VEGF-A121a to bind NRPs. Here, we review and attempt to reconcile evidence for and against VEGF-A121a binding to Neuropilins. This evidence suggests that, in vitro, VEGF-A121a can bind to both NRP1 and NRP2 via domains encoded by exons 5 and 8a; in the case of NRP1, VEGF-A121a binds with lower affinity than VEGF-A165a. In in vitro cell culture experiments, both NRP1 and NRP2 can enhance VEGF-A121a-induced phosphorylation of VEGFR2 and downstream signaling including proliferation. However, unlike VEGFA-165a, experiments have shown that VEGF-A121a does not ‘bridge’ VEGFR2 and NRP1, i.e. it does not bind both receptors simultaneously at their extracellular domain. Thus, the mechanism by which Neuropilins potentiate VEGF-A121a-mediated VEGFR2 signaling may be different from that for VEGF-A165a. We suggest such an alternate mechanism: interactions between NRP1 and VEGFR2 transmembrane (TM) and intracellular (IC) domains.

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“…Two regions within the receptor-binding domain of VEGF have been identified by charged-to-alanine scanning mutagenesis as being important determinants for KDR and Flt-l binding; positively charged residues close in sequence to LysW were found to be important for KDR binding, whereas negatively charged residues close in sequence to G1uW were associated with Flt-1 binding (Keyt et al, 1996b). In addition, introduction of a glycosylation site at residue 84 was shown to block binding to KDR (Keyt et al, 1996b).…”

mentioning

confidence: 99%

“…In addition, introduction of a glycosylation site at residue 84 was shown to block binding to KDR (Keyt et al, 1996b). A model of VEGF based upon the 3.0-A resolution crystal structure of PDGF-BB suggests that G~u~~ and LysS4 are distal in the monomer but proximal in the PDGF-like dimer (Keyt et al, 1996b).…”

mentioning

confidence: 99%

¹H, ¹³C, and ¹⁵N backbone assignment and secondary structure of the receptor‐binding domain of vascular endothelial growth factor

Fairbrother¹,

Champe²,

Christinger³

et al. 1997

Protein Science

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Nearly complete sequence-specific 'H, I3C, and I5N resonance assignments are reported for the backbone atoms of the receptor-binding domain of vascular endothelial growth factor (VEGF), a 23-kDa homodimeric protein that is a major regulator of both normal and pathological angiogenesis. The assignment strategy relied on the use of seven 3D triple-resonance experiments [HN(CO)CA, HNCA, HNCO, (HCA)CONH, HN(COCA)HA, HN(CA)HA, and CBCA-(CO)NH] and a 3D "N-TOCSY-HSQC experiment recorded on a 0.5 mM (12 mg/mL) sample at 500 MHz, pH 7.0, 45 "C. Under these conditions, I5N relaxation data show that the protein has a rotational correlation time of 15.0 ns. Despite this unusually long correlation time, assignments were obtained for 94 of the 99 residues; 8 residues lack amide 'H and I5N assignments, presumably due to rapid exchange of the amide 'H with solvent under the experimental conditions used. The secondary structure of the protein was deduced from the chemical shift indices of the 'H", "CU, I3Cp, and I3CO nuclei, and from analysis of backbone NOES observed in a 3D I5N-NOESY-HSQC spectrum. Two helices and a significant amount of P-sheet structure were identified, in general agreement with the secondary structure found in a recently determined crystal structure of a similar VEGF construct [Muller YA et al., 1997, Proc Natl Acad Sci USA 9 4 7 192-7 1971.

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Identification of Vascular Endothelial Growth Factor Determinants for Binding KDR and FLT-1 Receptors

Cited by 437 publications

References 58 publications

Roles of two VEGF receptors, Flt-1 and KDR, in the signal transduction of VEGF effects in human vascular endothelial cells

Roles of two VEGF receptors, Flt-1 and KDR, in the signal transduction of VEGF effects in human vascular endothelial cells

VEGF-A121a binding to Neuropilins – A concept revisited

¹H, ¹³C, and ¹⁵N backbone assignment and secondary structure of the receptor‐binding domain of vascular endothelial growth factor

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Identification of Vascular Endothelial Growth Factor Determinants for Binding KDR and FLT-1 Receptors

Cited by 437 publications

References 58 publications

Roles of two VEGF receptors, Flt-1 and KDR, in the signal transduction of VEGF effects in human vascular endothelial cells

Roles of two VEGF receptors, Flt-1 and KDR, in the signal transduction of VEGF effects in human vascular endothelial cells

VEGF-A121a binding to Neuropilins – A concept revisited

1H, 13C, and 15N backbone assignment and secondary structure of the receptor‐binding domain of vascular endothelial growth factor

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¹H, ¹³C, and ¹⁵N backbone assignment and secondary structure of the receptor‐binding domain of vascular endothelial growth factor