Fiche publication
Date publication
février 2017
Journal
Scientific reports
Auteurs
Membres identifiés du Cancéropôle Est :
Dr WAGNER Renaud
Tous les auteurs :
Chaptal V, Delolme F, Kilburg A, Magnard S, Montigny C, Picard M, Prier C, Monticelli L, Bornert O, Agez M, Ravaud S, Orelle C, Wagner R, Jawhari A, Broutin I, Pebay-Peyroula E, Jault JM, Kaback HR, le Maire M, Falson P
Lien Pubmed
Résumé
Most membrane proteins studies require the use of detergents, but because of the lack of a general, accurate and rapid method to quantify them, many uncertainties remain that hamper proper functional and structural data analyses. To solve this problem, we propose a method based on matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS) that allows quantification of pure or mixed detergents in complex with membrane proteins. We validated the method with a wide variety of detergents and membrane proteins. We automated the process, thereby allowing routine quantification for a broad spectrum of usage. As a first illustration, we show how to obtain information of the amount of detergent in complex with a membrane protein, essential for liposome or nanodiscs reconstitutions. Thanks to the method, we also show how to reliably and easily estimate the detergent corona diameter and select the smallest size, critical for favoring protein-protein contacts and triggering/promoting membrane protein crystallization, and to visualize the detergent belt for Cryo-EM studies.
Mots clés
Detergents, chemistry, Liposomes, Membrane Proteins, chemistry, Micelles, Models, Molecular, Reproducibility of Results, Sensitivity and Specificity, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, methods
Référence
Sci Rep. 2017 Feb 8;7:41751
