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

Abstract Most membrane proteins studies require the use of detergents, but because lack a general, accurate and rapid method to quantify them, many uncertainties remain that hamper proper functional structural data analyses. To solve this problem, we propose based on matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS) allows quantification pure or mixed detergents in complex with proteins. We validated wide variety automated process, thereby allowing routine for broad spectrum usage. As first illustration, show how obtain information amount detergent protein, essential liposome nanodiscs reconstitutions. Thanks method, also reliably easily estimate corona diameter select smallest size, critical favoring protein-protein contacts triggering/promoting protein crystallization, visualize belt Cryo-EM studies.