@article { ISI:000328682000016,
	title = {Mechanistic Insights from Substrate Preference in Unsaturated Glucuronyl Hydrolase},
	journal = {CHEMBIOCHEM},
	volume = {15},
	number = {1},
	year = {2014},
	month = {JAN 3},
	pages = {124-134},
	abstract = {Natural and synthetic unsaturated glucuronides were tested as substrates for Clostridium perfringens unsaturated glucuronyl hydrolase to probe its mechanism and to guide inhibitor design. Of the natural substrates, a chondroitin disaccharide substrate with sulfation of the primary alcohol on carbon 6 of its N-acetylgalactosamine moiety was found to have the highest turnover number of any substrate reported for an unsaturated glucuronyl hydrolase, with k(cat)=112 s(-1). Synthetic aryl glycoside substrates with electron-withdrawing aglycone substituents were cleaved more slowly than those with electron-donating substituents. Similarly, an unsaturated glucuronyl fluoride was found to be a particularly poor substrate, with k(cat)/K-m=44 nM(-1)s(-1)a very unusual result for a glycoside-cleaving enzyme. These results are consistent with a transition state with positive charge at carbon 5 and the endocyclic oxygen, as anticipated in the hydration mechanism proposed. However, several analogues designed to take advantage of strong enzyme binding to such a transition state showed little to no inhibition. This result suggests that further work is required to understand the true nature of the transition state stabilised by this enzyme.},
	issn = {1439-4227},
	doi = {10.1002/cbic.201300547},
	author = {Jongkees, Seino A. K. and Yoo, Hayoung and Withers, Stephen G.}
}
