Quantification of Detergents Complexed with Membrane Proteins

• DOI: 10.1038/srep41751
• Publication Date: 2017-02-08T11:55:39Z
• Authors: Vincent Chaptal; Frédéric Delolme; Arnaud Kilburg; Sandrine Magnard; Cédric Montigny; Martin Picard; Charlène Prier; Luca Monticelli; Olivier Bornert; Morgane Agez; Stéphanie Ravaud; Cédric Orelle; Renaud Wagner; Anass Jawhari; Isabelle Broutin; Eva Pebay-Peyroula; Jean-Michel Jault; H. Ronald Kaback; Marc le Maire; Pierre Falson
• Keywords: Models, Molecular; 0301 basic medicine; 570; [SDV]Life Sciences [q-bio]; Detergents; Aucun; [SDV.BBM.BP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics; Bioengineering; Sensitivity and Specificity; [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry; Article; Analytical Chemistry; 03 medical and health sciences; Models; 616; Matrix-Assisted Laser Desorption-Ionization; Micelles; 0303 health sciences; Molecular Biology/Structural Biology [q-bio.BM]; Spectrometry; Molecular; Membrane Proteins; Reproducibility of Results; [SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry; Biological Sciences; Mass; 540; [SDV] Life Sciences [q-bio]; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Chemical Sciences; Liposomes; Biochemistry and Cell Biology; Generic health relevance; Molecular Biology/Biophysics

AbstractMost 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.