5MI5

BtGH84 mutant with covalent modification by MA3 in complex with PUGNAc

Summary for 5MI5

• Entry DOI: 10.2210/pdb5mi5/pdb
• Descriptor: O-GlcNAcase BT_4395, O-(2-ACETAMIDO-2-DEOXY D-GLUCOPYRANOSYLIDENE) AMINO-N-PHENYLCARBAMATE, ~{N}-(4-ethoxyquinazolin-2-yl)propanamide, ... (5 entities in total)
• Functional Keywords: hydrolase, activator
• Biological source: Bacteroides thetaiotaomicron (strain ATCC 29148 / DSM 2079 / NCTC 10582 / E50 / VPI-5482)
• Total number of polymer chains: 1
• Total formula weight: 84240.13

Authors

Darby, J.F.,Davies, G.J.,Hubbard, R.E. (deposition date: 2016-11-27, release date: 2017-11-01, Last modification date: 2024-10-16)

Primary citation

Darby, J.F.,Atobe, M.,Firth, J.D.,Bond, P.,Davies, G.J.,O'Brien, P.,Hubbard, R.E.
Increase of enzyme activity through specific covalent modification with fragments.
Chem Sci, 8:7772-7779, 2017

PubMed 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 (/) 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.

PubMed: 29163914
DOI: 10.1039/c7sc01966a

Experimental method

X-RAY DIFFRACTION (2.15 Å)