Issue 4, 2020
From the journal:

RSC Chemical Biology

Long-range PEG stapling: macrocyclization for increased protein conformational stability and resistance to proteolysis†

Qiang Xiao,a   Dallin S. Ashton,a   Zachary B. Jones,a   Katherine P. Thompsona  and  Joshua L. Price ORCID logo *a  

* Corresponding authors

a Department of Chemistry and Biochemistry, Brigham Young University, Provo, Utah 84602, USA
E-mail: jlprice@chem.byu.edu

Abstract

We previously showed that long-range stapling of two Asn-linked O-allyl PEG oligomers via olefin metathesis substantially increases the conformational stability of the WW domain through an entropic effect. The impact of stapling was more favorable when the staple connected positions that were far apart in primary sequence but close in the folded tertiary structure. Here we validate these criteria by identifying new stabilizing PEG-stapling sites within the WW domain and the SH3 domain, both β-sheet proteins. We find that stapling via olefin metathesis vs. the copper(I)-catalyzed azide/alkyne cycloaddition (CuAAC) results in similar energetic benefits, suggesting that olefin and triazole staples can be used interchangeably. Proteolysis assays of selected WW variants reveal that the observed staple-based increases in conformational stability lead to enhanced proteolytic resistance. Finally, we find that an intermolecular staple dramatically increases the quaternary structural stability of an α-helical GCN4 coiled-coil heterodimer.

Graphical abstract: Long-range PEG stapling: macrocyclization for increased protein conformational stability and resistance to proteolysis

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Article information

DOI
https://doi.org/10.1039/D0CB00075B
Article type
Paper
Submitted
19 May 2020
Accepted
03 Aug 2020
First published
13 Aug 2020
This article is Open Access
Creative Commons BY-NC license

RSC Chem. Biol., 2020,1, 273-280

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Long-range PEG stapling: macrocyclization for increased protein conformational stability and resistance to proteolysis

Q. Xiao, D. S. Ashton, Z. B. Jones, K. P. Thompson and J. L. Price, RSC Chem. Biol., 2020, 1, 273 DOI: 10.1039/D0CB00075B

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