Characterization of two distinct modes of drug binding to human intestinal fatty acid binding protein

Rahul Patil, Aisha Khalidah Laguerre, Jerome Wielens, Stephen James Headey, Martin Williams, Maria Lourdes Regina Hughes, Biswaranjan Mohanty, Christopher John Porter, Martin Scanlon

Research output: Contribution to journalArticleResearchpeer-review

9 Citations (Scopus)

Abstract

The aqueous cytoplasm of cells poses a potentially significant barrier for many lipophilic drugs to reach their sites of action. Fatty acid binding proteins (FABPs) bind to poorly water-soluble fatty acids (FAs) and lipophilic compounds and facilitate their intracellular transport. Several structures of FA in complex with FABPs have been described, but data describing the binding sites of other lipophilic ligands including drugs are limited. Here the environmentally sensitive fluorophores, 1-anilinonapthalene 8-sulfonic acid (ANS), and 11-dansylamino undecanoic acid (DAUDA) were used to investigate drug binding to human intestinal FABP (hIFABP). Most drugs that bound hIFABP were able to displace both ANS and DAUDA. A notable exception was ketorolac, a non-steroidal anti-inflammatory drug that bound to hIFABP and displaced DAUDA but failed to displace ANS. Isothermal titration calorimetry revealed that for the majority of ligands including FA, ANS, and DAUDA, binding to hIFABP was exothermic. In contrast, ketorolac binding to hIFABP was endothermic and entropy-driven. The X-ray crystal structure of DAUDA-hIFABP revealed a FA-like binding mode where the carboxylate of DAUDA formed a network of hydrogen bonds with residues at the bottom of the binding cavity and the dansyl group interacted with residues in the portal region. In contrast, NMR chemical shift perturbation (CSP) data suggested that ANS bound only toward the bottom of the hIFABP cavity, whereas ketorolac occupied only the portal region. The CSP data further suggested that ANS and ketorolac were able to bind simultaneously to hIFABP, consistent with the lack of displacement of ANS observed by fluorescence and supported by a model of the ternary complex. The NMR solution structure of the ketorolac-hIFABP complex therefore describes a newly characterized, hydrophobic ligand binding site in the portal region of hIFABP.
Original languageEnglish
Pages (from-to)2526 - 2534
Number of pages9
JournalACS Chemical Biology
Volume9
Issue number11
DOIs
Publication statusPublished - 2014

Cite this

Patil, R., Laguerre, A. K., Wielens, J., Headey, S. J., Williams, M., Hughes, M. L. R., ... Scanlon, M. (2014). Characterization of two distinct modes of drug binding to human intestinal fatty acid binding protein. ACS Chemical Biology, 9(11), 2526 - 2534. https://doi.org/10.1021/cb5005178
Patil, Rahul ; Laguerre, Aisha Khalidah ; Wielens, Jerome ; Headey, Stephen James ; Williams, Martin ; Hughes, Maria Lourdes Regina ; Mohanty, Biswaranjan ; Porter, Christopher John ; Scanlon, Martin. / Characterization of two distinct modes of drug binding to human intestinal fatty acid binding protein. In: ACS Chemical Biology. 2014 ; Vol. 9, No. 11. pp. 2526 - 2534.
@article{bdb475906ebc4e5b883958573aabfbbb,
title = "Characterization of two distinct modes of drug binding to human intestinal fatty acid binding protein",
abstract = "The aqueous cytoplasm of cells poses a potentially significant barrier for many lipophilic drugs to reach their sites of action. Fatty acid binding proteins (FABPs) bind to poorly water-soluble fatty acids (FAs) and lipophilic compounds and facilitate their intracellular transport. Several structures of FA in complex with FABPs have been described, but data describing the binding sites of other lipophilic ligands including drugs are limited. Here the environmentally sensitive fluorophores, 1-anilinonapthalene 8-sulfonic acid (ANS), and 11-dansylamino undecanoic acid (DAUDA) were used to investigate drug binding to human intestinal FABP (hIFABP). Most drugs that bound hIFABP were able to displace both ANS and DAUDA. A notable exception was ketorolac, a non-steroidal anti-inflammatory drug that bound to hIFABP and displaced DAUDA but failed to displace ANS. Isothermal titration calorimetry revealed that for the majority of ligands including FA, ANS, and DAUDA, binding to hIFABP was exothermic. In contrast, ketorolac binding to hIFABP was endothermic and entropy-driven. The X-ray crystal structure of DAUDA-hIFABP revealed a FA-like binding mode where the carboxylate of DAUDA formed a network of hydrogen bonds with residues at the bottom of the binding cavity and the dansyl group interacted with residues in the portal region. In contrast, NMR chemical shift perturbation (CSP) data suggested that ANS bound only toward the bottom of the hIFABP cavity, whereas ketorolac occupied only the portal region. The CSP data further suggested that ANS and ketorolac were able to bind simultaneously to hIFABP, consistent with the lack of displacement of ANS observed by fluorescence and supported by a model of the ternary complex. The NMR solution structure of the ketorolac-hIFABP complex therefore describes a newly characterized, hydrophobic ligand binding site in the portal region of hIFABP.",
author = "Rahul Patil and Laguerre, {Aisha Khalidah} and Jerome Wielens and Headey, {Stephen James} and Martin Williams and Hughes, {Maria Lourdes Regina} and Biswaranjan Mohanty and Porter, {Christopher John} and Martin Scanlon",
year = "2014",
doi = "10.1021/cb5005178",
language = "English",
volume = "9",
pages = "2526 -- 2534",
journal = "ACS Chemical Biology",
issn = "1554-8929",
publisher = "American Chemical Society",
number = "11",

}

Characterization of two distinct modes of drug binding to human intestinal fatty acid binding protein. / Patil, Rahul; Laguerre, Aisha Khalidah; Wielens, Jerome; Headey, Stephen James; Williams, Martin; Hughes, Maria Lourdes Regina; Mohanty, Biswaranjan; Porter, Christopher John; Scanlon, Martin.

In: ACS Chemical Biology, Vol. 9, No. 11, 2014, p. 2526 - 2534.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Characterization of two distinct modes of drug binding to human intestinal fatty acid binding protein

AU - Patil, Rahul

AU - Laguerre, Aisha Khalidah

AU - Wielens, Jerome

AU - Headey, Stephen James

AU - Williams, Martin

AU - Hughes, Maria Lourdes Regina

AU - Mohanty, Biswaranjan

AU - Porter, Christopher John

AU - Scanlon, Martin

PY - 2014

Y1 - 2014

N2 - The aqueous cytoplasm of cells poses a potentially significant barrier for many lipophilic drugs to reach their sites of action. Fatty acid binding proteins (FABPs) bind to poorly water-soluble fatty acids (FAs) and lipophilic compounds and facilitate their intracellular transport. Several structures of FA in complex with FABPs have been described, but data describing the binding sites of other lipophilic ligands including drugs are limited. Here the environmentally sensitive fluorophores, 1-anilinonapthalene 8-sulfonic acid (ANS), and 11-dansylamino undecanoic acid (DAUDA) were used to investigate drug binding to human intestinal FABP (hIFABP). Most drugs that bound hIFABP were able to displace both ANS and DAUDA. A notable exception was ketorolac, a non-steroidal anti-inflammatory drug that bound to hIFABP and displaced DAUDA but failed to displace ANS. Isothermal titration calorimetry revealed that for the majority of ligands including FA, ANS, and DAUDA, binding to hIFABP was exothermic. In contrast, ketorolac binding to hIFABP was endothermic and entropy-driven. The X-ray crystal structure of DAUDA-hIFABP revealed a FA-like binding mode where the carboxylate of DAUDA formed a network of hydrogen bonds with residues at the bottom of the binding cavity and the dansyl group interacted with residues in the portal region. In contrast, NMR chemical shift perturbation (CSP) data suggested that ANS bound only toward the bottom of the hIFABP cavity, whereas ketorolac occupied only the portal region. The CSP data further suggested that ANS and ketorolac were able to bind simultaneously to hIFABP, consistent with the lack of displacement of ANS observed by fluorescence and supported by a model of the ternary complex. The NMR solution structure of the ketorolac-hIFABP complex therefore describes a newly characterized, hydrophobic ligand binding site in the portal region of hIFABP.

AB - The aqueous cytoplasm of cells poses a potentially significant barrier for many lipophilic drugs to reach their sites of action. Fatty acid binding proteins (FABPs) bind to poorly water-soluble fatty acids (FAs) and lipophilic compounds and facilitate their intracellular transport. Several structures of FA in complex with FABPs have been described, but data describing the binding sites of other lipophilic ligands including drugs are limited. Here the environmentally sensitive fluorophores, 1-anilinonapthalene 8-sulfonic acid (ANS), and 11-dansylamino undecanoic acid (DAUDA) were used to investigate drug binding to human intestinal FABP (hIFABP). Most drugs that bound hIFABP were able to displace both ANS and DAUDA. A notable exception was ketorolac, a non-steroidal anti-inflammatory drug that bound to hIFABP and displaced DAUDA but failed to displace ANS. Isothermal titration calorimetry revealed that for the majority of ligands including FA, ANS, and DAUDA, binding to hIFABP was exothermic. In contrast, ketorolac binding to hIFABP was endothermic and entropy-driven. The X-ray crystal structure of DAUDA-hIFABP revealed a FA-like binding mode where the carboxylate of DAUDA formed a network of hydrogen bonds with residues at the bottom of the binding cavity and the dansyl group interacted with residues in the portal region. In contrast, NMR chemical shift perturbation (CSP) data suggested that ANS bound only toward the bottom of the hIFABP cavity, whereas ketorolac occupied only the portal region. The CSP data further suggested that ANS and ketorolac were able to bind simultaneously to hIFABP, consistent with the lack of displacement of ANS observed by fluorescence and supported by a model of the ternary complex. The NMR solution structure of the ketorolac-hIFABP complex therefore describes a newly characterized, hydrophobic ligand binding site in the portal region of hIFABP.

UR - http://pubs.acs.org/doi/pdf/10.1021/cb5005178

U2 - 10.1021/cb5005178

DO - 10.1021/cb5005178

M3 - Article

VL - 9

SP - 2526

EP - 2534

JO - ACS Chemical Biology

JF - ACS Chemical Biology

SN - 1554-8929

IS - 11

ER -