Increase of enzyme activity through specific covalent modification with fragments

DOI: 10.1039/C7SC01966A

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)