期刊
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
卷 132, 期 16, 页码 5628-+出版社
AMER CHEMICAL SOC
DOI: 10.1021/ja9097498
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资金
- Russian Federal Agency for Science and Innovations
- Russian Federation
- Russian Academy of Science
- Russian Foundation for Basic Research
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 (similar to 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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