Identification and characterization of 5α-cyprinol-sulfating cytosolic sulfotransferases (Sults) in the zebrafish (Danio rerio)

Katsuhisa Kurogi; Maki Yoshihama; Austin Horton; Isaac T Schiefer; Matthew D Krasowski; Lee R Hagey; Frederick E Williams; Yoichi Sakakibara; Naoya Kenmochi; Masahito Suiko; Ming-Cheh Liu

doi:10.1016/j.jsbmb.2017.08.005

Identification and characterization of 5α-cyprinol-sulfating cytosolic sulfotransferases (Sults) in the zebrafish (Danio rerio)

J Steroid Biochem Mol Biol. 2017 Nov:174:120-127. doi: 10.1016/j.jsbmb.2017.08.005. Epub 2017 Aug 12.

Authors

Affiliations

¹ Department of Pharmacology, College of Pharmacy, University of Toledo Health Science Campus, Toledo, OH 43614, USA; Biochemistry and Applied Biosciences, University of Miyazaki, Miyazaki 889-2192, Japan.
² Department of Pharmacology, College of Pharmacy, University of Toledo Health Science Campus, Toledo, OH 43614, USA; Frontier Research Center, University of Miyazaki, Miyazaki 889-2192, Japan.
³ Department of Pharmacology, College of Pharmacy, University of Toledo Health Science Campus, Toledo, OH 43614, USA.
⁴ Department of Pathology, University of Iowa Hospitals and Clinics, RCP 6233, 200 Hawkins Drive, Iowa City, IA 52242, USA.
⁵ Department of Medicine, University of California at San Diego, La Jolla, San Diego, CA 92093, USA.
⁶ Biochemistry and Applied Biosciences, University of Miyazaki, Miyazaki 889-2192, Japan.
⁷ Frontier Research Center, University of Miyazaki, Miyazaki 889-2192, Japan.
⁸ Department of Pharmacology, College of Pharmacy, University of Toledo Health Science Campus, Toledo, OH 43614, USA. Electronic address: ming.liu@utoledo.edu.

Abstract

5α-Cyprinol 27-sulfate is the major biliary bile salt present in cypriniform fish including the zebrafish (Danio rerio). The current study was designed to identify the zebrafish cytosolic sulfotransferase (Sult) enzyme(s) capable of sulfating 5α-cyprinol and to characterize the zebrafish 5α-cyprinol-sulfating Sults in comparison with human SULT2A1. Enzymatic assays using zebrafish homogenates showed 5α-cyprinol-sulfating activity. A systematic analysis, using a panel of recombinant zebrafish Sults, revealed two Sult2 subfamily members, Sult2st2 and Sult2st3, as major 5α-cyprinol-sulfating Sults. Both enzymes showed higher activities using 5α-cyprinol as the substrate, compared to their activity with DHEA, a representative substrate for mammalian SULT2 family members, particularly SULT2A1. pH-Dependence and kinetics experiments indicated that the catalytic properties of zebrafish Sult2 family members in mediating the sulfation of 5α-cyprinol were different from those of either zebrafish Sult3st4 or human SULT2A1. Collectively, these results imply that both Sult2st2 and Sult2st3 have evolved to sulfate specifically C₂₇-bile alcohol, 5α-cyprinol, in Cypriniform fish, whereas the enzymatic characteristics of zebrafish Sult3 members, particularly Sult3st4, correlated with those of human SULT2A1.

Keywords: 5α-cyprinol; Cytosolic sulfotransferase; SULT; Sulfation; Zebrafish.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Animals
Arylsulfotransferase / metabolism*
Cholestanols / metabolism
Cholic Acids / metabolism
Dehydroepiandrosterone / metabolism
Embryo, Nonmammalian
Humans
Zebrafish
Zebrafish Proteins / metabolism*

Substances

Cholestanols
Cholic Acids
Zebrafish Proteins
anhydrocyprinol
Dehydroepiandrosterone
Arylsulfotransferase

Abstract

Publication types

MeSH terms

Substances

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