A click-flipped enzyme substrate boosts the performance of the diagnostic screening for Hunter syndrome.

Autor:	Schwarz M; Institute of Applied Synthetic Chemistry, TU Wien Getreidemarkt 9 1060 Vienna Austria hannes.mikula@tuwien.ac.at.; ARCHIMED Life Science GmbH Leberstraße 20 1110 Vienna Austria., Skrinjar P; Institute of Applied Synthetic Chemistry, TU Wien Getreidemarkt 9 1060 Vienna Austria hannes.mikula@tuwien.ac.at., Fink MJ; Department of Chemistry and Chemical Biology, Harvard University 12 Oxford Street Cambridge MA 02138 USA., Kronister S; Institute of Applied Synthetic Chemistry, TU Wien Getreidemarkt 9 1060 Vienna Austria hannes.mikula@tuwien.ac.at., Mechtler T; ARCHIMED Life Science GmbH Leberstraße 20 1110 Vienna Austria., Koukos PI; Bijvoet Centre for Biomolecular Research, Faculty of Science - Chemistry, Utrecht University Padualaan 8 3584CH Utrecht The Netherlands., Bonvin AMJJ; Bijvoet Centre for Biomolecular Research, Faculty of Science - Chemistry, Utrecht University Padualaan 8 3584CH Utrecht The Netherlands., Kasper DC; ARCHIMED Life Science GmbH Leberstraße 20 1110 Vienna Austria., Mikula H; Institute of Applied Synthetic Chemistry, TU Wien Getreidemarkt 9 1060 Vienna Austria hannes.mikula@tuwien.ac.at.
Jazyk:	angličtina
Zdroj:	Chemical science [Chem Sci] 2020 Oct 23; Vol. 11 (47), pp. 12671-12676. Date of Electronic Publication: 2020 Oct 23.
DOI:	10.1039/d0sc04696e
Abstrakt:	We report on the unexpected finding that click modification of iduronyl azides results in a conformational flip of the pyranose ring, which led to the development of a new strategy for the design of superior enzyme substrates for the diagnostic assaying of iduronate-2-sulfatase (I2S), a lysosomal enzyme related to Hunter syndrome. Synthetic substrates are essential in testing newborns for metabolic disorders to enable early initiation of therapy. Our click-flipped iduronyl triazole showed a remarkably better performance with I2S than commonly used O -iduronates. We found that both O - and triazole-linked substrates are accepted by the enzyme, irrespective of their different conformations, but only the O -linked product inhibits the activity of I2S. Thus, in the long reaction times required for clinical assays, the triazole substrate substantially outperforms the O -iduronate. Applying our click-flipped substrate to assay I2S in dried blood spots sampled from affected patients and random newborns significantly increased the confidence in discriminating between these groups, clearly indicating the potential of the click-flip strategy to control the biomolecular function of carbohydrates. Competing Interests: There are no conflicts to declare. (This journal is © The Royal Society of Chemistry.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu