Unlabeled lysophosphatidic acid receptor binding in free solution as determined by a compensated interferometric reader.

Journal of Lipid Research
Manisha RayJerold Chun

Abstract

Native interactions between lysophospholipids (LPs) and their cognate LP receptors are difficult to measure because of lipophilicity and/or the adhesive properties of lipids, which contribute to high levels of nonspecific binding in cell membrane preparations. Here, we report development of a free-solution assay (FSA) where label-free LPs bind to their cognate G protein-coupled receptors (GPCRs), combined with a recently reported compensated interferometric reader (CIR) to quantify native binding interactions between receptors and ligands. As a test case, the binding parameters between lysophosphatidic acid (LPA) receptor 1 (LPA1; one of six cognate LPA GPCRs) and LPA were determined. FSA-CIR detected specific binding through the simultaneous real-time comparison of bound versus unbound species by measuring the change in the solution dipole moment produced by binding-induced conformational and/or hydration changes. FSA-CIR identified KD values for chemically distinct LPA species binding to human LPA1 and required only a few nanograms of protein: 1-oleoyl (18:1; KD = 2.08 ± 1.32 nM), 1-linoleoyl (18:2; KD = 2.83 ± 1.64 nM), 1-arachidonoyl (20:4; KD = 2.59 ± 0.481 nM), and 1-palmitoyl (16:0; KD = 1.69 ± 0.1 nM) LPA. These KD valu...Continue Reading

References

Feb 13, 2002·Drug Discovery Today·Alan WiseStephen Rees
Jun 19, 2002·Biochimica Et Biophysica Acta·Amy H YangJerold Chun
Jun 3, 2004·Chemistry and Physics of Lipids·Zaiguo LiRobert Bittman
Oct 29, 2005·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Lutea A A de JongRainer Bischoff
Jan 30, 2007·Biophysical Journal·Jordi Faraudo, Alex Travesset
Apr 24, 2009·The Journal of Biological Chemistry·Keisuke YanagidaSatoshi Ishii
Jan 9, 2010·Annual Review of Pharmacology and Toxicology·Ji Woong ChoiJerold Chun
Oct 29, 2010·Critical Reviews in Biochemistry and Molecular Biology·Kyoko Noguchi, Jerold Chun
Dec 24, 2010·Proceedings of the National Academy of Sciences of the United States of America·Ji Woong ChoiJerold Chun
Mar 15, 2011·Nature Biotechnology·Michael M BakshDarryl J Bornhop
Apr 27, 2011·Annals of Neurology·Jeffrey A Cohen, Jerold Chun
Jun 28, 2011·Methods in Molecular Biology·Kimiko Kuroki, Katsumi Maenaka
Jul 19, 2011·ACS Medicinal Chemistry Letters·Iva NavratilovaAndrew L Hopkins
Aug 16, 2011·British Journal of Pharmacology·Tetsuji MutohJerold Chun
Jan 28, 2014·Biomedical Spectroscopy and Imaging·Silvia Locatelli-HoopsInna Gorshkova
Mar 8, 2014·British Journal of Pharmacology·Yasuyuki KiharaJerold Chun
Mar 20, 2014·Journal of Lipid Research·Yun C YungJerold Chun
Jul 11, 2014·ACS Chemical Biology·Francisco CiruelaVíctor Fernández-Dueñas
Sep 18, 2014·The Analyst·Michael N KammerDarryl J Bornhop
Dec 17, 2014·Experimental Cell Research·Yasuyuki KiharaJerold Chun
Jan 17, 2015·Biomolecules & Therapeutics·Nicole C Stoddard, Jerold Chun
Feb 20, 2015·Neuron·Yun C YungJerold Chun
Apr 9, 2015·Development·Xiaoyan ShengJerold Chun
Jun 20, 2015·Cell·Jill E ChrencikMichael A Hanson
Sep 1, 2015·British Journal of Pharmacology·Leigh A StoddartKevin D G Pfleger
Oct 29, 2015·Nature Reviews. Molecular Cell Biology·Antoine-Emmanuel SalibaAnne-Claude Gavin
Nov 29, 2017·Trends in Pharmacological Sciences·Michael Kauk, Carsten Hoffmann
Feb 6, 2018·Optics Letters·Michael N KammerDarryl J Bornhop
May 29, 2018·European Journal of Pharmacology·Jonathan ElleryMatt Barnes
Jul 10, 2018·ACS Sensors·Michael N KammerDarryl J Bornhop
Oct 23, 2018·Trends in Pharmacological Sciences·Victoria A Blaho, Jerold Chun
Jan 10, 2019·Annual Review of Pharmacology and Toxicology·Jerold ChunVictoria A Blaho
Apr 26, 2019·ACS Combinatorial Science·Michael N KammerDarryl J Bornhop
Jul 18, 2019·Analytical Chemistry·Michael N KammerDarryl J Bornhop

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Citations

Nov 6, 2020·Neuromolecular Medicine·Eric Birgbauer
Jan 6, 2022·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Yasuyuki KiharaJerold Chun

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Methods Mentioned

BETA
surface plasmon resonance
dynamic light scattering
interaction studies

Software Mentioned

GraphPad
CIR
FSA

Related Concepts

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ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.

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