A ligand-induced structural change in fatty acid-binding protein 1 is associated with potentiation of peroxisome proliferator-activated receptor α agonists

Rahul Patil, Biswaranjan Mohanty, Bonan Liu, Indu R Chandrashekaran, Stephen J Headey, Martin L Williams, Craig S Clements, Olga Ilyichova, Bradley C Doak, Patrick Genissel, Richard Weaver, Laurent Vuillard, Michelle L Halls, Christopher J H Porter, Martin J Scanlon

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Peroxisome proliferator-activated receptor α (PPARα) is a transcriptional regulator of lipid metabolism. GW7647 is a potent PPARα agonist that must reach the nucleus to activate this receptor. In cells expressing human fatty acid-binding protein 1 (FABP1), GW7647 treatment increases FABP1's nuclear localization and potentiates GW7647-mediated PPARα activation; GW7647 is less effective in cells that do not express FABP1. To elucidate the underlying mechanism, here we substituted residues in FABP1 known to dictate lipid signaling by other intracellular lipid-binding proteins. Substitutions of Lys-20 and Lys-31 to Ala in the FABP1 helical cap affected neither its nuclear localization nor PPARα activation. In contrast, Ala substitution of Lys-57, Glu-77, and Lys-96, located in the loops adjacent to the ligand-binding portal region, abolished both FABP1 nuclear localization and GW7647-induced PPARα activation but had little effect on GW7647-FABP1 binding affinity. Using solution NMR spectroscopy, we determined the WT FABP1 structure and analyzed the dynamics in the apo and GW7647-bound structures of both the WT and the K57A/E77A/K96A triple mutant. We found that GW7647 binding causes little change in the FABP1 backbone, but solvent exposes several residues in the loops around the portal region, including Lys-57, Glu-77, and Lys-96. These residues also become more solvent-exposed upon binding of FABP1 with the endogenous PPARα agonist oleic acid. Together with previous observations, our findings suggest that GW7647 binding stabilizes a FABP1 conformation that promotes its interaction with PPARα. We conclude that full PPARα agonist activity of GW7647 requires FABP1-dependent transport and nuclear localization processes.

Original languageEnglish
Pages (from-to)3720-3734
Number of pages15
JournalJournal of Biological Chemistry
Volume294
Issue number10
DOIs
Publication statusPublished - 8 Mar 2019

Keywords

  • drug delivery
  • fatty acid–binding protein
  • GW7647
  • nuclear magnetic resonance (NMR)
  • nuclear receptor
  • peroxisome proliferator-activated receptor (PPAR)
  • PPARα agonist
  • protein structure
  • transcription factor

Cite this

@article{5b4298fd5a74449290ada8bdf80cd214,
title = "A ligand-induced structural change in fatty acid-binding protein 1 is associated with potentiation of peroxisome proliferator-activated receptor α agonists",
abstract = "Peroxisome proliferator-activated receptor α (PPARα) is a transcriptional regulator of lipid metabolism. GW7647 is a potent PPARα agonist that must reach the nucleus to activate this receptor. In cells expressing human fatty acid-binding protein 1 (FABP1), GW7647 treatment increases FABP1's nuclear localization and potentiates GW7647-mediated PPARα activation; GW7647 is less effective in cells that do not express FABP1. To elucidate the underlying mechanism, here we substituted residues in FABP1 known to dictate lipid signaling by other intracellular lipid-binding proteins. Substitutions of Lys-20 and Lys-31 to Ala in the FABP1 helical cap affected neither its nuclear localization nor PPARα activation. In contrast, Ala substitution of Lys-57, Glu-77, and Lys-96, located in the loops adjacent to the ligand-binding portal region, abolished both FABP1 nuclear localization and GW7647-induced PPARα activation but had little effect on GW7647-FABP1 binding affinity. Using solution NMR spectroscopy, we determined the WT FABP1 structure and analyzed the dynamics in the apo and GW7647-bound structures of both the WT and the K57A/E77A/K96A triple mutant. We found that GW7647 binding causes little change in the FABP1 backbone, but solvent exposes several residues in the loops around the portal region, including Lys-57, Glu-77, and Lys-96. These residues also become more solvent-exposed upon binding of FABP1 with the endogenous PPARα agonist oleic acid. Together with previous observations, our findings suggest that GW7647 binding stabilizes a FABP1 conformation that promotes its interaction with PPARα. We conclude that full PPARα agonist activity of GW7647 requires FABP1-dependent transport and nuclear localization processes.",
keywords = "drug delivery, fatty acid–binding protein, GW7647, nuclear magnetic resonance (NMR), nuclear receptor, peroxisome proliferator-activated receptor (PPAR), PPARα agonist, protein structure, transcription factor",
author = "Rahul Patil and Biswaranjan Mohanty and Bonan Liu and Chandrashekaran, {Indu R} and Headey, {Stephen J} and Williams, {Martin L} and Clements, {Craig S} and Olga Ilyichova and Doak, {Bradley C} and Patrick Genissel and Richard Weaver and Laurent Vuillard and Halls, {Michelle L} and Porter, {Christopher J H} and Scanlon, {Martin J}",
year = "2019",
month = "3",
day = "8",
doi = "10.1074/jbc.RA118.006848",
language = "English",
volume = "294",
pages = "3720--3734",
journal = "Journal of Biological Chemistry",
issn = "1083-351X",
publisher = "American Society for Biochemistry and Molecular Biology",
number = "10",

}

A ligand-induced structural change in fatty acid-binding protein 1 is associated with potentiation of peroxisome proliferator-activated receptor α agonists. / Patil, Rahul ; Mohanty, Biswaranjan; Liu, Bonan; Chandrashekaran, Indu R; Headey, Stephen J; Williams, Martin L; Clements, Craig S; Ilyichova, Olga; Doak, Bradley C; Genissel, Patrick; Weaver, Richard; Vuillard, Laurent; Halls, Michelle L; Porter, Christopher J H; Scanlon, Martin J.

In: Journal of Biological Chemistry, Vol. 294, No. 10, 08.03.2019, p. 3720-3734.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - A ligand-induced structural change in fatty acid-binding protein 1 is associated with potentiation of peroxisome proliferator-activated receptor α agonists

AU - Patil, Rahul

AU - Mohanty, Biswaranjan

AU - Liu, Bonan

AU - Chandrashekaran, Indu R

AU - Headey, Stephen J

AU - Williams, Martin L

AU - Clements, Craig S

AU - Ilyichova, Olga

AU - Doak, Bradley C

AU - Genissel, Patrick

AU - Weaver, Richard

AU - Vuillard, Laurent

AU - Halls, Michelle L

AU - Porter, Christopher J H

AU - Scanlon, Martin J

PY - 2019/3/8

Y1 - 2019/3/8

N2 - Peroxisome proliferator-activated receptor α (PPARα) is a transcriptional regulator of lipid metabolism. GW7647 is a potent PPARα agonist that must reach the nucleus to activate this receptor. In cells expressing human fatty acid-binding protein 1 (FABP1), GW7647 treatment increases FABP1's nuclear localization and potentiates GW7647-mediated PPARα activation; GW7647 is less effective in cells that do not express FABP1. To elucidate the underlying mechanism, here we substituted residues in FABP1 known to dictate lipid signaling by other intracellular lipid-binding proteins. Substitutions of Lys-20 and Lys-31 to Ala in the FABP1 helical cap affected neither its nuclear localization nor PPARα activation. In contrast, Ala substitution of Lys-57, Glu-77, and Lys-96, located in the loops adjacent to the ligand-binding portal region, abolished both FABP1 nuclear localization and GW7647-induced PPARα activation but had little effect on GW7647-FABP1 binding affinity. Using solution NMR spectroscopy, we determined the WT FABP1 structure and analyzed the dynamics in the apo and GW7647-bound structures of both the WT and the K57A/E77A/K96A triple mutant. We found that GW7647 binding causes little change in the FABP1 backbone, but solvent exposes several residues in the loops around the portal region, including Lys-57, Glu-77, and Lys-96. These residues also become more solvent-exposed upon binding of FABP1 with the endogenous PPARα agonist oleic acid. Together with previous observations, our findings suggest that GW7647 binding stabilizes a FABP1 conformation that promotes its interaction with PPARα. We conclude that full PPARα agonist activity of GW7647 requires FABP1-dependent transport and nuclear localization processes.

AB - Peroxisome proliferator-activated receptor α (PPARα) is a transcriptional regulator of lipid metabolism. GW7647 is a potent PPARα agonist that must reach the nucleus to activate this receptor. In cells expressing human fatty acid-binding protein 1 (FABP1), GW7647 treatment increases FABP1's nuclear localization and potentiates GW7647-mediated PPARα activation; GW7647 is less effective in cells that do not express FABP1. To elucidate the underlying mechanism, here we substituted residues in FABP1 known to dictate lipid signaling by other intracellular lipid-binding proteins. Substitutions of Lys-20 and Lys-31 to Ala in the FABP1 helical cap affected neither its nuclear localization nor PPARα activation. In contrast, Ala substitution of Lys-57, Glu-77, and Lys-96, located in the loops adjacent to the ligand-binding portal region, abolished both FABP1 nuclear localization and GW7647-induced PPARα activation but had little effect on GW7647-FABP1 binding affinity. Using solution NMR spectroscopy, we determined the WT FABP1 structure and analyzed the dynamics in the apo and GW7647-bound structures of both the WT and the K57A/E77A/K96A triple mutant. We found that GW7647 binding causes little change in the FABP1 backbone, but solvent exposes several residues in the loops around the portal region, including Lys-57, Glu-77, and Lys-96. These residues also become more solvent-exposed upon binding of FABP1 with the endogenous PPARα agonist oleic acid. Together with previous observations, our findings suggest that GW7647 binding stabilizes a FABP1 conformation that promotes its interaction with PPARα. We conclude that full PPARα agonist activity of GW7647 requires FABP1-dependent transport and nuclear localization processes.

KW - drug delivery

KW - fatty acid–binding protein

KW - GW7647

KW - nuclear magnetic resonance (NMR)

KW - nuclear receptor

KW - peroxisome proliferator-activated receptor (PPAR)

KW - PPARα agonist

KW - protein structure

KW - transcription factor

UR - http://www.scopus.com/inward/record.url?scp=85062633455&partnerID=8YFLogxK

U2 - 10.1074/jbc.RA118.006848

DO - 10.1074/jbc.RA118.006848

M3 - Article

VL - 294

SP - 3720

EP - 3734

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 1083-351X

IS - 10

ER -