The structure of the catechin-binding site of human sulfotransferase 1A1

Ian T. Cook, Ting Wang, Mark Girvin, Thomas S. Leyh

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

We are just beginning to understand the allosteric regulation of the human cytosolic sulfotransferase (SULTs) family-13 disease-relevant enzymes that regulate the activities of hundreds, if not thousands, of signaling small molecules. SULT1A1, the predominant isoform in adult liver, harbors two noninteracting allosteric sites, each of which binds a differentmolecular family: The catechins (naturally occurring flavonols) and nonsteroidal antiinflammatory drugs (NSAIDs). Here, we present the structure of an SULT allosteric binding site-the catechin-binding site of SULT1A1 bound to epigallocatechin gallate (EGCG). The allosteric pocket resides in a dynamic region of the protein that enables EGCG to control opening and closure of the enzyme's active-site cap. Furthermore, the structure offers a molecular explanation for the isozyme specificity of EGCG, which is corroborated experimentally. The bindingsite structure was obtained without X-ray crystallography or multidimensional NMR. Instead, a SULT1A1 apoprotein structure was used to guide positioning of a small number of spin-labeled single-Cysmutants that coat the entire enzyme surface with a paramagnetic field of sufficient strength to determine its contribution to the bound ligand's transverse (T2) relaxation from its 1D solution spectrum. EGCG protons were mapped to the protein surface by triangulation using the T2 values to calculate their distances to a trio of spin-labeled Cys mutants. The final structure was obtained using distance-constrained molecular dynamics docking. This approach, which is readily extensible to other systems, is applicable over a wide range of ligand affinities, requires little protein, avoids the need for isotopically labeled protein, and has no protein molecular weight limitations.

Original languageEnglish (US)
Pages (from-to)14312-14317
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume113
Issue number50
DOIs
StatePublished - Dec 13 2016

Fingerprint

Catechin
Binding Sites
Allosteric Site
Sulfotransferases
Proteins
Enzymes
Allosteric Regulation
Ligands
Flavonols
Apoproteins
X Ray Crystallography
Molecular Dynamics Simulation
Isoenzymes
Protons
Catalytic Domain
Protein Isoforms
Membrane Proteins
Anti-Inflammatory Agents
Molecular Weight
epigallocatechin gallate

Keywords

  • Allostery
  • Catechin
  • Nmr
  • Structure
  • Sulfotransferase

ASJC Scopus subject areas

  • General

Cite this

The structure of the catechin-binding site of human sulfotransferase 1A1. / Cook, Ian T.; Wang, Ting; Girvin, Mark; Leyh, Thomas S.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 113, No. 50, 13.12.2016, p. 14312-14317.

Research output: Contribution to journalArticle

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