Quantification of Detergents Complexed with Membrane Proteins

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
DOI: 10.1038/srep41751 Publication Date: 2017-02-08T11:55:39Z
ABSTRACT
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.
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