Difference between revisions of "Part:BBa K5302000"

Latest revision as of 08:11, 1 October 2024

VEGFR1D2

VEGFR1D2: VEGF binds to two tyrosine kinase receptors, VEGF receptor 1 (VEGFR1/Flt-1) and receptor 2 (VEGFR2/KDR), which are localized on the cell surface of various cell types. The receptors are organized in two intracellular kinase domains, a short transmembrane region and an extracellular portion constituted of seven immunoglobulin-like domains. The binding of VEGF to the extracellular domain induces receptor dimerization, transphosphorylation of the kinase domains, and activation of the intracellular signaling. Deletion studies have shown that the ligand binding function resides within the first three domains of VEGFR14 and in domains 2 and 3 of VEGFR2. Deletion of the second extracellular domain of VEGFR1 (VEGFR1D2) completely abolishes the binding to VEGF, whereas VEGFR1D2 binds VEGF with an affinity that is only 60 times lower than the entire extracellular portion of the receptor. The equilibrium constant for the binding of VEGF to one VEGFR1D2 molecule was determined by surface plasmon resonance. The obtained value, 2.9 ± 0.2 nM, is consistent with the constant reported for the binding of VEGF to monomeric VEGFR1D2. We used pBBR plasmid as a backbone and transfered VEGFR1D2 into Escherichia coli Nissle 1917, and finally succeeded in VEGFR1D2-expressing.

Figure 1. Predictions of VEGFR1D2 Secondary Structure

Figure 2. VEGFR1D2 Trypsin Digestion Analysis(Numbers refer to the human VEGFR1 protein sequence, underlined residues are involved in a disulfide bridge, the proline in bold indicates that the Arg-Pro bond is not cleaved by trypsin22. exp = experimental, calc = calculated.)

Figure 3. structure of VEGFR1D2

Figure 4. Colony PCR results of pBBR-OmpA-VEGFR1D2

Figure 5. Colony PCR results of pBBR-INP-VEGFR1D2 (1)

Figure 6. Colony PCR results of pBBR-INP-VEGFR1D2 (2)

Figure 7. Colony PCR results of pBBR-INP-VEGFR1D2 (3)

Sequence and Features