Functional display of bioactive peptides on the vGFP scaffold

Sharon Min Qi Chee, Jantana Wongsantichon, Lau Sze Yi, Barindra Sana, Yuri Frosi, Robert C. Robinson, Farid J. Ghadessy

Research output: Contribution to journalArticlepeer-review


Grafting bioactive peptides into recipient protein scaffolds can often increase their activities by conferring enhanced stability and cellular longevity. Here, we describe use of vGFP as a novel scaffold to display peptides. vGFP comprises GFP fused to a bound high affinity Enhancer nanobody that potentiates its fluorescence. We show that peptides inserted into the linker region between GFP and the Enhancer are correctly displayed for on-target interaction, both in vitro and in live cells by pull-down, measurement of target inhibition and imaging analyses. This is further confirmed by structural studies highlighting the optimal display of a vGFP-displayed peptide bound to Mdm2, the key negative regulator of p53 that is often overexpressed in cancer. We also demonstrate a potential biosensing application of the vGFP scaffold by showing target-dependent modulation of intrinsic fluorescence. vGFP is relatively thermostable, well-expressed and inherently fluorescent. These properties make it a useful scaffold to add to the existing tool box for displaying peptides that can disrupt clinically relevant protein–protein interactions.

Original languageEnglish
Article number10127
JournalScientific reports
Issue number1
Publication statusPublished - Dec 2021

ASJC Scopus subject areas

  • General


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