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
AUTHORS (20)
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.
SUPPLEMENTAL MATERIAL
Coming soon ....
REFERENCES (58)
CITATIONS (64)
EXTERNAL LINKS
PlumX Metrics
RECOMMENDATIONS
FAIR ASSESSMENT
Coming soon ....
JUPYTER LAB
Coming soon ....