Human DHEA sulfation requires direct interaction between PAPS synthase 2 and DHEA sulfotransferase SULT2A1
File(s)
Published version
Author(s)
Type
Journal Article
Abstract
The high-energy sulfate donor 3′-phosphoadenosine-5′-phosphosulfate (PAPS), generated by human PAPS synthase isoforms PAPSS1 and PAPSS2, is required for all human sulfation pathways. Sulfotransferase SULT2A1 uses PAPS for sulfation of the androgen precursor dehydroepiandrosterone (DHEA), thereby reducing downstream activation of DHEA to active androgens. Human PAPSS2 mutations manifest with undetectable DHEA sulfate, androgen excess, and metabolic disease, suggesting that ubiquitous PAPSS1 cannot compensate for deficient PAPSS2 in supporting DHEA sulfation. In knockdown studies in human adrenocortical NCI-H295R1 cells, we found that PAPSS2, but not PAPSS1, is required for efficient DHEA sulfation. Specific APS kinase activity, the rate-limiting step in PAPS biosynthesis, did not differ between PAPSS1 and PAPSS2. Co-expression of cytoplasmic SULT2A1 with a cytoplasmic PAPSS2 variant supported DHEA sulfation more efficiently than co-expression with nuclear PAPSS2 or nuclear/cytosolic PAPSS1. Proximity ligation assays revealed protein–protein interactions between SULT2A1 and PAPSS2 and, to a lesser extent, PAPSS1. Molecular docking studies showed a putative binding site for SULT2A1 within the PAPSS2 APS kinase domain. Energy-dependent scoring of docking solutions identified the interaction as specific for the PAPSS2 and SULT2A1 isoforms. These findings elucidate the mechanistic basis for the selective requirement for PAPSS2 in human DHEA sulfation.
Date Issued
2018-06-22
Date Acceptance
2018-04-28
Citation
Journal of Biological Chemistry, 2018, 293 (25), pp.9724-9735
ISSN
0021-9258
Publisher
Elsevier
Start Page
9724
End Page
9735
Journal / Book Title
Journal of Biological Chemistry
Volume
293
Issue
25
Copyright Statement
© 2018 Mueller et al. Published by The American Society for Biochemistry and Molecular Biology, Inc. This is an Open Access article under the CC BY license
License URL
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/29743239
PII: S0021-9258(20)39958-0
Subjects
Biochemistry & Molecular Biology
CRYSTAL-STRUCTURE
CYTOSOLIC SULFOTRANSFERASES
dehydroepiandrosterone
DHEAS
DOMAIN
enzyme kinetics
ENZYMES
EVOLUTION
EXPRESSION
Life Sciences & Biomedicine
LOCALIZATION
molecular docking
MUTATIONS
PAPS synthase
PROTEIN-PROTEIN DOCKING
protein-protein interaction
QUINARY STRUCTURE
Science & Technology
steroid hormone
steroid sulfation pathway
sulfotransferase
Publication Status
Published
Coverage Spatial
United States
Date Publish Online
2018-05-09