Key result
Lipid-protein nanodiscs guide detergent selection to preserve native VSD structure for NMR studies.
Why the study?
The choice of a suitable detergent-based membrane mimetic is crucial for high-resolution NMR studies of membrane proteins, but optimal screening methods are needed.
Lipid-protein nanodiscs can serve as an effective reference medium for screening detergents to preserve the native structure of integral membrane proteins in NMR studies.
LPN reference spectra may optimize detergent choice for native-like VSD NMR samples; leaves open translation to functional or in vivo cardiac ion channel studies.
The choice of a suitable detergent-based membrane mimetic is of crucial importance for high-resolution NMR studies of membrane proteins. The present report describes a new approach of detergent screening. It is based on the comparison of 2D (1)H,(15)N-correlation spectra of a protein in a membrane-bilayer "reference" medium and in "trial" detergent-based environments. The proposed "reference" medium is the lipid-protein nanodisc (LPN) representing nanoscale phospholipid bilayers wrapped around by apolipoprotein A-1. The set of zwitterionic (DPC, DMPC/DHPC), anionic (SDS, LMPG, LPPG), and weakly cationic (LDAO) detergent-based media was screened for their ability to represent the native structure of the isolated voltage-sensing domain (VSD) of the archaeal potassium channel KvAP. The VSD/LPN complexes composed of saturated zwitterionic (DMPC), anionic (DMPG), or a mixture of unsaturated differently charged (POPC/DOPG, 3:1) lipids were used as reference. All assayed detergent media demonstrate similar CD spectra of the domain with a high level (approximately 60%) of overall helicity but different 2D NMR spectra. Using the reference spectrum of the VSD in LPN, we were able to choose the detergent composition in which the membrane-like structure of the VSD is preserved.
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Шенкарев et al. (2010) studied this question. Lipid-protein nanodiscs (LPN) vs. Detergent-based media was evaluated on Preservation of the membrane-like structure of the voltage-sensing domain (VSD) in NMR spectra. Lipid-protein nanodiscs used as a reference medium enabled the selection of a detergent composition that preserves the membrane-like structure of the voltage-sensing domain for NMR studies.
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