Functional display of bioactive peptides on the vGFP scaffold

Abstract 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 nanobod...

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Autores principales:	Sharon Min Qi Chee, Jantana Wongsantichon, Lau Sze Yi, Barindra Sana, Yuri Frosi, Robert C. Robinson, Farid J. Ghadessy
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/bb66fff127b041c1baa2825939769763
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spelling	oai:doaj.org-article:bb66fff127b041c1baa28259397697632021-12-02T16:50:32ZFunctional display of bioactive peptides on the vGFP scaffold10.1038/s41598-021-89421-y2045-2322https://doaj.org/article/bb66fff127b041c1baa28259397697632021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-89421-yhttps://doaj.org/toc/2045-2322Abstract 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.Sharon Min Qi CheeJantana WongsantichonLau Sze YiBarindra SanaYuri FrosiRobert C. RobinsonFarid J. GhadessyNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sharon Min Qi Chee Jantana Wongsantichon Lau Sze Yi Barindra Sana Yuri Frosi Robert C. Robinson Farid J. Ghadessy Functional display of bioactive peptides on the vGFP scaffold
description	Abstract 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.
format	article
author	Sharon Min Qi Chee Jantana Wongsantichon Lau Sze Yi Barindra Sana Yuri Frosi Robert C. Robinson Farid J. Ghadessy
author_facet	Sharon Min Qi Chee Jantana Wongsantichon Lau Sze Yi Barindra Sana Yuri Frosi Robert C. Robinson Farid J. Ghadessy
author_sort	Sharon Min Qi Chee
title	Functional display of bioactive peptides on the vGFP scaffold
title_short	Functional display of bioactive peptides on the vGFP scaffold
title_full	Functional display of bioactive peptides on the vGFP scaffold
title_fullStr	Functional display of bioactive peptides on the vGFP scaffold
title_full_unstemmed	Functional display of bioactive peptides on the vGFP scaffold
title_sort	functional display of bioactive peptides on the vgfp scaffold
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/bb66fff127b041c1baa2825939769763
work_keys_str_mv	AT sharonminqichee functionaldisplayofbioactivepeptidesonthevgfpscaffold AT jantanawongsantichon functionaldisplayofbioactivepeptidesonthevgfpscaffold AT lauszeyi functionaldisplayofbioactivepeptidesonthevgfpscaffold AT barindrasana functionaldisplayofbioactivepeptidesonthevgfpscaffold AT yurifrosi functionaldisplayofbioactivepeptidesonthevgfpscaffold AT robertcrobinson functionaldisplayofbioactivepeptidesonthevgfpscaffold AT faridjghadessy functionaldisplayofbioactivepeptidesonthevgfpscaffold
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Functional display of bioactive peptides on the vGFP scaffold

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