Abstract
The ability to design detergents that are suitable for protein analysis by mass spectrometry (MS) represents an on-going challenge in the field of native MS. Desirable detergent characteristics include charge-reducing properties and low gas-phase stabilities of complexes formed with proteins. In this work, the gas-phase properties of oligoglycerol detergents (OGDs) are optimized by fine tuning of their molecular structure. Furthermore, a tandem mass spectrometry (MS/MS) approach is presented that estimates the gas-phase properties of detergents simply by studying the dissociation behaviour of protein-detergent complexes (PDCs) formed with the soluble protein β-lactoglobulin (BLG).Graphical ᅟ
Highlights
Detergents are commonly applied for the structural investigation of membrane proteins, which relies on the fact that they are used traditionally for membrane protein purification
To evaluate the impact of individual detergent building blocks on the gas-phase properties of dendritic oligoglycerol detergents (OGDs), three soluble proteins were assessed as model systems for the investigation of protein-detergent complexes (PDCs): two hydrophilic proteins, ubiquitin (8.5 kDa), and myoglobin (16.9 kDa) which are known to not bind detergents in solution under native conditions, and β-lactoglobulin (BLG, 18.4 kDa), a more amphiphilic protein that is suggested to play an important role in the transport of amphiphilic molecules [18]. In line with this suggestion, we found that BLG exhibits a greater propensity to form PDCs upon nanoelectrospray ionization (nESI) than ubiquitin or myoglobin
The mixtures were transferred into the gas phase by nESI from ammonium acetate buffer (10 mM) under instrumental conditions chosen to optimize the intensity of PDC signals
Summary
Detergents are commonly applied for the structural investigation of membrane proteins, which relies on the fact that they are used traditionally for membrane protein purification. Membrane proteins that are encapsulated in detergent aggregates are usually transferred into the gas phase of a mass spectrometer by means of nanoelectrospray ionization (nESI), where the excess of detergent is subsequently removed via thermal activation [2, 3]. The activation conditions required for detergent removal and the charge state of the released protein are important parameters for the preservation of compact protein structures. The structural diversity of available detergent families makes it challenging to deduce design principles for detergents that allow for on-demand adjustment of gas-phase properties. To overcome this limitation, we here investigated the gas-phase
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