Solid-state nuclear magnetic resonance evidence for an extended beta strand conformation of the membrane-bound HIV-1 fusion peptide.
Solid-state nuclear magnetic resonance (NMR) spectroscopy was applied to the membrane-bound form of a synthetic peptide representing the 23-residue N-terminal fusion peptide domain of the HIV-1 gp41 envelope glycoprotein. 1D solid-state NMR line width measurements of singly 13C carbonyl labeled peptides showed that a significant population of the membrane-bound peptide is well-structured in its N-terminal and central regions while the C-terminus has more disorder. There was some dependence of line width on lipid composition, with narrower line widths and hence greater structural order observed for a lipid composition comparable to that found in the virus and its target T cells. In the more ordered N-terminal and central regions of the peptide, the 13C carbonyl chemical shifts are consistent with a nonhelical membrane-bound conformation. Additional evidence for a beta strand membrane-bound conformation was provided by analysis of 2D rotor-synchronized magic angle spinning NMR spectra of doubly 13C carbonyl labeled peptides. Lipid mixing and aqueous contents leakage assays were applied to demonstrate the fusogenicity of the peptide under conditions comparable to those used for the solid-state NMR sample preparation.
Yang
J
J
Department of Chemistry, Michigan State University, East Lansing, Michigan 48824, USA.
Gabrys
C M
CM
Weliky
D P
DP
D000595:Amino Acid Sequence; D002244:Carbon / Q000737:chemistry; D015700:HIV Envelope Protein gp41 / Q000737:chemistry* / Q000378:metabolism; D015497:HIV-1 / Q000737:chemistry*; D008051:Lipid Bilayers / Q000378:metabolism; D008561:Membrane Fusion; D008563:Membrane Lipids / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D010446:Peptide Fragments / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D012996:Solutions; D014760:Viral Fusion Proteins / Q000737:chemistry* / Q000378:metabolism; D014867:Water
D015700:HIV Envelope Protein gp41; D008051:Lipid Bilayers; D008563:Membrane Lipids; D010446:Peptide Fragments; D012996:Solutions; D014760:Viral Fusion Proteins; D014867:Water; D002244:Carbon