Screening ligands for membrane protein receptors by total internal reflection fluorescence: the 5-HT3 serotonin receptor

Analytical Chemistry
E L SchmidH Vogel

Abstract

The screening of ligands for membrane receptor proteins is central to the discovery of new pharmaceutical drugs. We present a general method to reversibly attach receptor proteins via an affinity tag to a quartz surface and subsequently detect with high sensitivity the real-time binding of ligands by total internal reflection fluorescence. A serotonin-gated ion channel protein was immobilized, and the binding of a fluorescent ligand was investigated. The affinity and the kinetic parameters of binding were measured, and the effect of unlabeled compounds was determined by competition. The pharmacology of the immobilized receptor was identical to that of the native receptor. The affinity of unlabeled ligands was rapidly and effectively determined. The method described here is generally applicable for membrane proteins and opens new ways for the discovery of pharmacologically active compounds.

References

Nov 1, 1989·Analytical Biochemistry·K R RogersM E Eldefrawi
Jun 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·J L YakelR A North
Jan 1, 1996·Biosensors & Bioelectronics·T E PlowmanJ N Herron

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Citations

Dec 18, 2003·Journal of Applied Microbiology·K E SapsfordF S Ligler
Apr 15, 2014·The Journal of Membrane Biology·Eduardo Antonio Della PiaKaren L Martinez
Mar 11, 1999·Journal of Receptor and Signal Transduction Research·R HoviusH Vogel
Jan 1, 2009·Proceedings of the National Academy of Sciences of the United States of America·Delphine CharvolinJean-Luc Popot
Mar 11, 2004·The European Journal of Neuroscience·Régis GrailhePierre-Jean Corringer
Oct 6, 2000·Methods : a Companion to Methods in Enzymology·W M MullettJ M Yeung
Jul 6, 2000·Analytical Chemistry·P B OldhamI M Warner
Mar 19, 2021·Chemistry : a European Journal·Carlotta BorgarelliErmal Ismalaj
Apr 28, 2005·Biosensors & Bioelectronics·Chris Rowe TaittFrances S Ligler
Nov 28, 2012·ACS Chemical Neuroscience·Evgenia G MatveevaRichard B Thompson
Sep 8, 2004·Langmuir : the ACS Journal of Surfaces and Colloids·Per RiglerHorst Vogel

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