Allosteres to regulate neurotransmitter sulfonation

Kristie Darrah, Ting Wang, Ian T. Cook, Mary Cacace, Alexander Deiters, Thomas S. Leyh

Research output: Contribution to journalArticle

Abstract

Catecholamine neurotransmitter levels in the synapses of the brain shape human disposition— cognitive flexibility, aggression, depression, and reward seeking—and manipulating these levels is a major objective of the pharmaceutical industry. Certain neurotransmitters are extensively sulfonated and inactivated by human sulfotransferase 1A3 (SULT1A3). To our knowledge, sulfonation as a therapeutic means of regulating transmitter activity has not been explored. Here, we describe the discovery of a SULT1A3 allosteric site that can be used to inhibit the enzyme. The structure of the new site is determined using spin-label-triangulation NMR. The site forms a cleft at the edge of a conserved 30-residue active-site cap that must open and close during the catalytic cycle. Allosteres anchor into the site via -stacking interactions with two residues that sandwich the planar core of the allostere and inhibit the enzyme through cap-stabilizing interactions with substituents attached to the core. Changes in cap free energy were calculated ab initio as a function of core substituents and used to design and synthesize a series of inhibitors intended to progressively stabilize the cap and slow turnover. The inhibitors bound tightly (34 nM to 7.4 M) and exhibited progressive inhibition. The cap-stabilizing effects of the inhibitors were experimentally determined and agreed remarkably well with the theoretical predictions. These studies establish a reliable heuristic for the design of SULT1A3 allosteric inhibitors and demonstrate that the free-energy changes of a small, dynamic loop that is critical for SULT substrate selection and turnover can be calculated accurately.

Original languageEnglish (US)
Pages (from-to)2293-2301
Number of pages9
JournalJournal of Biological Chemistry
Volume294
Issue number7
DOIs
StatePublished - Jan 1 2019

Fingerprint

Sulfotransferases
Sulfonation
Neurotransmitter Agents
Free energy
Allosteric Site
Spin Labels
Drug Industry
Triangulation
Enzymes
Anchors
Aggression
Reward
Synapses
Catecholamines
Transmitters
Brain
Catalytic Domain
Nuclear magnetic resonance
Depression
Substrates

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Allosteres to regulate neurotransmitter sulfonation. / Darrah, Kristie; Wang, Ting; Cook, Ian T.; Cacace, Mary; Deiters, Alexander; Leyh, Thomas S.

In: Journal of Biological Chemistry, Vol. 294, No. 7, 01.01.2019, p. 2293-2301.

Research output: Contribution to journalArticle

Darrah, Kristie ; Wang, Ting ; Cook, Ian T. ; Cacace, Mary ; Deiters, Alexander ; Leyh, Thomas S. / Allosteres to regulate neurotransmitter sulfonation. In: Journal of Biological Chemistry. 2019 ; Vol. 294, No. 7. pp. 2293-2301.
@article{e6dc93a8d3f7405c965a04d2ed69f6cf,
title = "Allosteres to regulate neurotransmitter sulfonation",
abstract = "Catecholamine neurotransmitter levels in the synapses of the brain shape human disposition— cognitive flexibility, aggression, depression, and reward seeking—and manipulating these levels is a major objective of the pharmaceutical industry. Certain neurotransmitters are extensively sulfonated and inactivated by human sulfotransferase 1A3 (SULT1A3). To our knowledge, sulfonation as a therapeutic means of regulating transmitter activity has not been explored. Here, we describe the discovery of a SULT1A3 allosteric site that can be used to inhibit the enzyme. The structure of the new site is determined using spin-label-triangulation NMR. The site forms a cleft at the edge of a conserved 30-residue active-site cap that must open and close during the catalytic cycle. Allosteres anchor into the site via -stacking interactions with two residues that sandwich the planar core of the allostere and inhibit the enzyme through cap-stabilizing interactions with substituents attached to the core. Changes in cap free energy were calculated ab initio as a function of core substituents and used to design and synthesize a series of inhibitors intended to progressively stabilize the cap and slow turnover. The inhibitors bound tightly (34 nM to 7.4 M) and exhibited progressive inhibition. The cap-stabilizing effects of the inhibitors were experimentally determined and agreed remarkably well with the theoretical predictions. These studies establish a reliable heuristic for the design of SULT1A3 allosteric inhibitors and demonstrate that the free-energy changes of a small, dynamic loop that is critical for SULT substrate selection and turnover can be calculated accurately.",
author = "Kristie Darrah and Ting Wang and Cook, {Ian T.} and Mary Cacace and Alexander Deiters and Leyh, {Thomas S.}",
year = "2019",
month = "1",
day = "1",
doi = "10.1074/jbc.RA118.006511",
language = "English (US)",
volume = "294",
pages = "2293--2301",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "7",

}

TY - JOUR

T1 - Allosteres to regulate neurotransmitter sulfonation

AU - Darrah, Kristie

AU - Wang, Ting

AU - Cook, Ian T.

AU - Cacace, Mary

AU - Deiters, Alexander

AU - Leyh, Thomas S.

PY - 2019/1/1

Y1 - 2019/1/1

N2 - Catecholamine neurotransmitter levels in the synapses of the brain shape human disposition— cognitive flexibility, aggression, depression, and reward seeking—and manipulating these levels is a major objective of the pharmaceutical industry. Certain neurotransmitters are extensively sulfonated and inactivated by human sulfotransferase 1A3 (SULT1A3). To our knowledge, sulfonation as a therapeutic means of regulating transmitter activity has not been explored. Here, we describe the discovery of a SULT1A3 allosteric site that can be used to inhibit the enzyme. The structure of the new site is determined using spin-label-triangulation NMR. The site forms a cleft at the edge of a conserved 30-residue active-site cap that must open and close during the catalytic cycle. Allosteres anchor into the site via -stacking interactions with two residues that sandwich the planar core of the allostere and inhibit the enzyme through cap-stabilizing interactions with substituents attached to the core. Changes in cap free energy were calculated ab initio as a function of core substituents and used to design and synthesize a series of inhibitors intended to progressively stabilize the cap and slow turnover. The inhibitors bound tightly (34 nM to 7.4 M) and exhibited progressive inhibition. The cap-stabilizing effects of the inhibitors were experimentally determined and agreed remarkably well with the theoretical predictions. These studies establish a reliable heuristic for the design of SULT1A3 allosteric inhibitors and demonstrate that the free-energy changes of a small, dynamic loop that is critical for SULT substrate selection and turnover can be calculated accurately.

AB - Catecholamine neurotransmitter levels in the synapses of the brain shape human disposition— cognitive flexibility, aggression, depression, and reward seeking—and manipulating these levels is a major objective of the pharmaceutical industry. Certain neurotransmitters are extensively sulfonated and inactivated by human sulfotransferase 1A3 (SULT1A3). To our knowledge, sulfonation as a therapeutic means of regulating transmitter activity has not been explored. Here, we describe the discovery of a SULT1A3 allosteric site that can be used to inhibit the enzyme. The structure of the new site is determined using spin-label-triangulation NMR. The site forms a cleft at the edge of a conserved 30-residue active-site cap that must open and close during the catalytic cycle. Allosteres anchor into the site via -stacking interactions with two residues that sandwich the planar core of the allostere and inhibit the enzyme through cap-stabilizing interactions with substituents attached to the core. Changes in cap free energy were calculated ab initio as a function of core substituents and used to design and synthesize a series of inhibitors intended to progressively stabilize the cap and slow turnover. The inhibitors bound tightly (34 nM to 7.4 M) and exhibited progressive inhibition. The cap-stabilizing effects of the inhibitors were experimentally determined and agreed remarkably well with the theoretical predictions. These studies establish a reliable heuristic for the design of SULT1A3 allosteric inhibitors and demonstrate that the free-energy changes of a small, dynamic loop that is critical for SULT substrate selection and turnover can be calculated accurately.

UR - http://www.scopus.com/inward/record.url?scp=85061593300&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85061593300&partnerID=8YFLogxK

U2 - 10.1074/jbc.RA118.006511

DO - 10.1074/jbc.RA118.006511

M3 - Article

VL - 294

SP - 2293

EP - 2301

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 7

ER -