Darby, John F. orcid.org/0000-0003-2754-6348, Atobe, Masakazu, Firth, James D. et al. (4 more authors) (2017) Increase of enzyme activity through specific covalent modification with fragments. Chemical Science. pp. 7772-7779. ISSN 2041-6539
Abstract
Modulation of enzyme activity is a powerful means of probing cellular function and can be exploited for diverse applications. Here, we explore a method of enzyme activation where covalent tethering of a small molecule to an enzyme can increase catalytic activity (k cat/K M) up to 35-fold. Using a bacterial glycoside hydrolase, BtGH84, we demonstrate how small molecule "fragments", identified as activators in free solution, can be covalently tethered to the protein using Michael-addition chemistry. We show how tethering generates a constitutively-activated enzyme-fragment conjugate, which displays both improved catalytic efficiency and increased susceptibility to certain inhibitor classes. Structure guided modifications of the tethered fragment demonstrate how specific interactions between the fragment and the enzyme influence the extent of activation. This work suggests that a similar approach may be used to modulate the activity of enzymes such as to improve catalytic efficiency or increase inhibitor susceptibility.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © The Royal Society of Chemistry 2017. |
Dates: |
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Institution: | The University of York |
Academic Units: | The University of York > Faculty of Sciences (York) > Chemistry (York) |
Funding Information: | Funder Grant number BBSRC (BIOTECHNOLOGY AND BIOLOGICAL SCIENCES RESEARCH COUNCIL) BB/N008332/1 |
Depositing User: | Pure (York) |
Date Deposited: | 27 Sep 2017 14:15 |
Last Modified: | 16 Oct 2024 14:04 |
Published Version: | https://doi.org/10.1039/C7SC01966A |
Status: | Published |
Refereed: | Yes |
Identification Number: | 10.1039/C7SC01966A |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:121760 |