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Functional Expression of All Human Sulfotransferases in Fission Yeast, Assay Development, and Structural Models for Isoforms SULT4A1 and SULT6B1

1
School of Pharmaceutical Science and Technology, Health Sciences Platform, Tianjin University, Tianjin 300072, China
2
Pharmaceutical and Medicinal Chemistry (Pharmaceutical Analyses), Institute of Pharmacy, Freie Universitaet Berlin, 14195 Berlin, Germany
3
Pharmaceutical and Medicinal Chemistry (Computer-Aided Drug Design), Institute of Pharmacy, Freie Universitaet Berlin, 14195 Berlin, Germany
*
Authors to whom correspondence should be addressed.
Biomolecules 2020, 10(11), 1517; https://doi.org/10.3390/biom10111517
Received: 12 October 2020 / Revised: 3 November 2020 / Accepted: 4 November 2020 / Published: 6 November 2020
Cytosolic sulfotransferases (SULTs) catalyze phase II (conjugation) reactions of drugs and endogenous compounds. A complete set of recombinant fission yeast strains each expressing one of the 14 human SULTs was generated, including SULT4A1 and SULT6B1. Sulfation of test substrates by whole-cell biotransformation was successfully demonstrated for all enzymes for which substrates were previously known. The results proved that the intracellular production of the cofactor 3′-phosphoadenosine 5′-phosphosulfate (PAPS) necessary for SULT activity in fission yeast is sufficiently high to support metabolite production. A modified variant of sulfotransferase assay was also developed that employs permeabilized fission yeast cells (enzyme bags). Using this approach, SULT4A1-dependent sulfation of 1-naphthol was observed. Additionally, a new and convenient SULT activity assay is presented. It is based on the sulfation of a proluciferin compound, which was catalyzed by SULT1E1, SULT2A1, SULT4A1, and SULT6B1. For the latter two enzymes this study represents the first demonstration of their enzymatic functionality. Furthermore, the first catalytically competent homology models for SULT4A1 and SULT6B1 in complex with PAPS are reported. Through mechanistic molecular modeling driven by substrate docking, we pinned down the increased activity levels of these two isoforms to optimized substrate binding. View Full-Text
Keywords: drug metabolism; phase II; proluciferin; sulfation; sulfotransferase drug metabolism; phase II; proluciferin; sulfation; sulfotransferase
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MDPI and ACS Style

Sun, Y.; Machalz, D.; Wolber, G.; Parr, M.K.; Bureik, M. Functional Expression of All Human Sulfotransferases in Fission Yeast, Assay Development, and Structural Models for Isoforms SULT4A1 and SULT6B1. Biomolecules 2020, 10, 1517. https://doi.org/10.3390/biom10111517

AMA Style

Sun Y, Machalz D, Wolber G, Parr MK, Bureik M. Functional Expression of All Human Sulfotransferases in Fission Yeast, Assay Development, and Structural Models for Isoforms SULT4A1 and SULT6B1. Biomolecules. 2020; 10(11):1517. https://doi.org/10.3390/biom10111517

Chicago/Turabian Style

Sun, Yanan, David Machalz, Gerhard Wolber, Maria Kristina Parr, and Matthias Bureik. 2020. "Functional Expression of All Human Sulfotransferases in Fission Yeast, Assay Development, and Structural Models for Isoforms SULT4A1 and SULT6B1" Biomolecules 10, no. 11: 1517. https://doi.org/10.3390/biom10111517

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