A general method to quantify ligand-driven oligomerization from fluorescence-based images

Stoneman, M. R., Biener, G., Ward, R. J., Pediani, J. D. , Badu, D., Eis, A., Popa, I., Milligan, G. and Raicu, V. (2019) A general method to quantify ligand-driven oligomerization from fluorescence-based images. Nature Methods, 16(6), pp. 493-496. (doi: 10.1038/s41592-019-0408-9) (PMID:31110281)

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Abstract

Here, we introduce fluorescence intensity fluctuation spectrometry for determining the identity, abundance and stability of protein oligomers. This approach was tested on monomers and oligomers of known sizes and was used to uncover the oligomeric states of the epidermal growth factor receptor and the secretin receptor in the presence and absence of their agonist ligands. This method is fast and is scalable for high-throughput screening of drugs targeting protein–protein interactions.

Item Type:Articles
Additional Information:This work was partly funded by the National Science Foundation grant PHY-1626450 (awarded to V.R.), the Medical Research Council (U.K.) grant MR/L023806/1 (to G.M.), and the UWM Research Growth Initiative grants 101X333 (to V.R.) and 101X340 (to I.P.).
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Pediani, Dr John and Ward, Dr Richard and Milligan, Professor Graeme and Raicu, Professor Valerica
Authors: Stoneman, M. R., Biener, G., Ward, R. J., Pediani, J. D., Badu, D., Eis, A., Popa, I., Milligan, G., and Raicu, V.
College/School:College of Medical Veterinary and Life Sciences > School of Molecular Biosciences
Journal Name:Nature Methods
Publisher:Nature Research
ISSN:1548-7091
ISSN (Online):1548-7105
Published Online:20 May 2019
Copyright Holders:Copyright © 2019 Springer Nature
First Published:First published in Nature Methods 16(6): 493-496
Publisher Policy:Reproduced in accordance with the copyright policy of the publisher

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
656901The organisational structure of class A GPCRs: Implications for pharmacology, function and therapeutic regulationGraeme MilliganMedical Research Council (MRC)MR/L023806/1RI MOLECULAR CELL & SYSTEMS BIOLOGY