Quantification of Detergents Complexed with Membrane Proteins.
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.
Laboratory of Molecular Microbiology and Structural Biochemistry, CNRS UMR 5086, IBCP, Lyon 69367 France.
Vincent
V
Chaptal
Protein Science Facility, UMS 3444, IBCP, Lyon 69367, France.
Frédéric
F
Delolme
Laboratory of Molecular Microbiology and Structural Biochemistry, CNRS UMR 5086, IBCP, Lyon 69367 France.
Arnaud
A
Kilburg
Laboratory of Molecular Microbiology and Structural Biochemistry, CNRS UMR 5086, IBCP, Lyon 69367 France.
Sandrine
S
Magnard
Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, University of Paris-Sud, 91198 Gif-sur-Yvette, France.
Cédric
C
Montigny
Laboratoire de Biologie Physico-Chimique des Protéines Membranaires, CNRS UMR 7099, 75005 Paris, France.
Martin
M
Picard
Laboratory of Molecular Microbiology and Structural Biochemistry, CNRS UMR 5086, IBCP, Lyon 69367 France.
Charlène
C
Prier
Laboratory of Molecular Microbiology and Structural Biochemistry, CNRS UMR 5086, IBCP, Lyon 69367 France.
Luca
L
Monticelli
Institut de Recherche de l'Ecole de Biotechnologie de Strasbourg, University of Strasbourg-CNRS, Illkirch, France.
Olivier
O
Bornert
CALIXAR, 60 Avenue Rockefeller - 69008 Lyon, France.
Morgane
M
Agez
Institut de Biologie Structurale (IBS), University of Grenoble Alpes, CEA, CNRS, 38044 Grenoble, France.
Stéphanie
S
Ravaud
Laboratory of Molecular Microbiology and Structural Biochemistry, CNRS UMR 5086, IBCP, Lyon 69367 France.
Cédric
C
Orelle
Institut de Recherche de l'Ecole de Biotechnologie de Strasbourg, University of Strasbourg-CNRS, Illkirch, France.
Renaud
R
Wagner
CALIXAR, 60 Avenue Rockefeller - 69008 Lyon, France.
Anass
A
Jawhari
Laboratoire de Cristallographie et RMN Biologiques, CNRS UMR 8015, 75006 Paris, France.
Isabelle
I
Broutin
Institut de Biologie Structurale (IBS), University of Grenoble Alpes, CEA, CNRS, 38044 Grenoble, France.
Eva
E
Pebay-Peyroula
Laboratory of Molecular Microbiology and Structural Biochemistry, CNRS UMR 5086, IBCP, Lyon 69367 France.
Jean-Michel
JM
Jault
Department of Physiology, David Geffen School of Medicine, UCLA, Los Angeles, CA 90095 USA.
H Ronald
HR
Kaback
Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, University of Paris-Sud, 91198 Gif-sur-Yvette, France.
Marc
M
le Maire
Laboratory of Molecular Microbiology and Structural Biochemistry, CNRS UMR 5086, IBCP, Lyon 69367 France.
Pierre
P
Falson
D003902:Detergents / Q000737:chemistry* / Q000378:metabolism; D008081:Liposomes; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D008823:Micelles; D008958:Models, Molecular; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods / Q000592:standards
D003902:Detergents; D008081:Liposomes; D008565:Membrane Proteins; D008823:Micelles