Abstract
Diacyl phosphatidylglycerols (PG) are the major anionic lipids in the Escherichia coli membrane. Short-chain dihexanoyl phosphatidylglycerol (DHPG) was previously utilized for the structural determination, by NMR spectroscopy, of the peptide corresponding to the N-terminal membrane anchor of the glucose-specific enzyme IIA (IIAGlc) from E. coli. This study explores the possible use of lipid micelles of dioctanoyl phosphatidylglycerol (DOPG) and didecanoyl phosphatidylglycerol (DDPG) as alternatives to DHPG. At a peptide concentration of 1 mM, the minimum peptide/lipid molar ratios required for the formation of the lipid-binding amphipathic helix are approximately 1:40, 1:5, and 1:5 for DHPG, DOPG, and DDPG, respectively. Based on the lipid titration, the critical micelle concentration (CMC) of DHPG was estimated to be ∼50 mM. The 1H spectral linewidths of the peptide bound to a variety of lipid micelles decrease in the following order: DDPG > DOPG > DHPG. The helical regions of the peptide in different anionic lipids were elucidated based on chemical shift indexes (CSI). Residues Leu2-Leu9, Leu2-Val10, and Leu2-Val10 were found to be helical in DHPG, DOPG, and DDPG, respectively, indicating that the lipid chain length had only a subtle effect on the amphipathic helix of the peptide. In light of the minimum peptide/lipid ratio and the spectral linewidth, and the CSI-derived peptide structure, DOPG is proposed as a good compromise for structural studies of this membrane-associated peptide by solution NMR spectroscopy.
Original language | English (US) |
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Pages (from-to) | 257-264 |
Number of pages | 8 |
Journal | Spectroscopy |
Volume | 18 |
Issue number | 2 |
DOIs | |
State | Published - 2004 |
Keywords
- Amphipathic helix
- CMC
- Escherichia coli
- Lipid chain length
- Micelles
- NMR
ASJC Scopus subject areas
- Spectroscopy