Fragment-derived modulators of an industrial β-glucosidase.

Autor:	Makraki E; Department of Chemistry, University of York, Heslington, York YO10 5DD, U.K., Darby JF; Department of Chemistry, University of York, Heslington, York YO10 5DD, U.K.; Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield Medical School, Beech Hill, Sheffield, U.K., Carneiro MG; ZoBio BV, J.H. Oortweg 19, 2333 CH Leiden, The Netherlands., Firth JD; Department of Chemistry, University of York, Heslington, York YO10 5DD, U.K., Heyam A; Department of Chemistry, University of York, Heslington, York YO10 5DD, U.K., Ab E; ZoBio BV, J.H. Oortweg 19, 2333 CH Leiden, The Netherlands., O'Brien P; Department of Chemistry, University of York, Heslington, York YO10 5DD, U.K., Siegal G; ZoBio BV, J.H. Oortweg 19, 2333 CH Leiden, The Netherlands.; Division of Medicinal Chemistry, Amsterdam Institute for Molecules, Medicines and Systems (AIMMS), Vrije Universiteit, De Boelelaan 1108, 1081 HZ Amsterdam, Netherlands., Hubbard RE; Department of Chemistry, University of York, Heslington, York YO10 5DD, U.K.; Vernalis Research, Granta Park, Abington, Cambridge CB21 6GB, U.K.
Jazyk:	angličtina
Zdroj:	The Biochemical journal [Biochem J] 2020 Nov 27; Vol. 477 (22), pp. 4383-4395.
DOI:	10.1042/BCJ20200507
Abstrakt:	A fragment screen of a library of 560 commercially available fragments using a kinetic assay identified a small molecule that increased the activity of the fungal glycoside hydrolase TrBgl2. An analogue by catalogue approach and detailed kinetic analysis identified improved compounds that behaved as nonessential activators with up to a 2-fold increase in maximum activation. The compounds did not activate the related bacterial glycoside hydrolase CcBglA demonstrating specificity. Interestingly, an analogue of the initial fragment inhibits both TrBgl2 and CcBglA, apparently through a mixed-model mechanism. Although it was not possible to determine crystal structures of activator binding to 55 kDa TrBgl2, solution NMR experiments demonstrated a specific binding site for the activator. A partial assignment of the NMR spectrum gave the identity of the amino acids at this site, allowing a model for TrBgl2 activation to be built. The activator binds at the entrance of the substrate-binding site, generating a productive conformation for the enzyme-substrate complex. (© 2020 The Author(s).)
Databáze:	MEDLINE
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