Effect of the conformation of a peptide from gp41 on binding and domain formation in model membranes

Abstract

Binding of the peptide fragment 828–84. (P828), amino acid sequence RVIEVVQGACRAIRHIPRRIR, from the carboxy-terminal region of the envelope glycoprotein gp41 of human immunodeficiency virus type 1 (HIV-1) to membranes composed of a mixture of neutral and negatively charged phospholipids results in domain or cluster formation of the charged lipid. The conformation and dynamics of the peptide are investigated in solution and in the presence of sodium dodecyl sulphate (SDS) micelles using high resolution nuclear magnetic resonance (NMR) spectroscopy and circular dichroism (CD) spectropolarimetry. The CD results demonstrate that addition of either SDS, negatively charged phospholipid liposomes, or trifluoroethanol (TFE) induces a conformational transition of the peptide from a random coil or an extended chain in water to a more ordered structure with an estimated helical content of up to 60%. The structure of the peptide in a membrane mimetic SDS solution was investigated in detail using two-dimensional NMR. The measurements demonstrate the existence of a helical component in the peptide conformation in the SDS-bound state. The peptide most likely exists as an ensemble of conformations with exchange times between them which are fast on the chemical shift NMR time scale (10^-3 s). Simple neutralization of the six arginine sidechain charges does not cause the peptide to adopt an ordered structure. Thus, there is an additional requirement for the structural transition such as that resulting from constraint of the peptide on a surface, or localization of the peptide at the lipid-water interface where the polarity is lower. Our results favour a model of the peptide-lipid interaction in which the peptide backbone is located in the water phase and part of the amino acid sidechains penetrate the lipid-water interface.

Key Words:

• Conformation
• dynamic equilibrium
• peptide-lipid interaction
• NMR