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Increase of enzyme activity through specific covalent modification with fragments
Authors:
John F.
Darby
(University of York)
,
Masakazu
Atobe
(University of York; Asahi Kasei Pharma Corporation)
,
James D.
Firth
(University of York)
,
Paul
Bond
(University of York)
,
Gideon J.
Davies
(University of York)
,
Peter
O'Brien
(University of York)
,
Roderick E.
Hubbard
(University of York; Vernalis (R&D) Ltd)
Co-authored by industrial partner:
No
Type:
Journal Paper
Journal:
Chemical Science
, VOL 8
, PAGES 7772 - 7779
State:
Published (Approved)
Published:
September 2017

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 (kcat/KM) 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.
Diamond Keywords: Enzymes
Subject Areas:
Chemistry,
Biology and Bio-materials
Instruments:
I02-Macromolecular Crystallography
,
I03-Macromolecular Crystallography
Added On:
02/11/2017 15:58
Discipline Tags:
Biotechnology
Biochemistry
Catalysis
Chemistry
Structural biology
Engineering & Technology
Life Sciences & Biotech
Technical Tags:
Diffraction
Macromolecular Crystallography (MX)