Difference between revisions of "Part:BBa K5302003"

Latest revision as of 02:59, 2 October 2024

ZVEGF

This year, the USTC iGEM team has utilized the competitive binding of vascular endothelial growth factor (VEGF) to develop a targeted bacterial therapy for solid tumors. Our quest for the optimal VEGF-binding protein(or peptide) led us to an in-depth exploration of proteins structurally akin to the vascular endothelial growth factor receptor (VEGFR), which we have named VEGFR-like. ZVEGF is a 59-residue three-helix peptide that was developed by Fedorova et al. by randomizing 9 residues on helices 1 and 2 of the Z-domain scaffold via phage display, followed by selection for binding to the VEGF8-109 dimer. A co-crystal structure of ZVEGF with VEGF8-109 shows that the engineered Z-domain adopts the expected three-helix bundle tertiary structure and engages the receptor-recognition sites on the VEGF dimer through a surface formed by helices 1 and 2 of ZVEGF. It shows great affinity with VEGF(Ki=0.41 μM).We used pBBR1MCS-2 plasmid as a backbone and transfered ZVEGF into Escherichia coli Nissle 1917, and finally succeeded in expressing ZVEGF.

Figure 1. ZVEGF sequence

Figure 2. VEGF-binding site of ZVEGF

Figure 3. Colony PCR results of pBBR-OmpA-ZVEGF

Sequence and Features