Expression and biochemical analysis of the entire HIV-2 gp41 ectodomain: determinants of stability map to N- and C-terminal sequences outside the 6-helix bundle core

FEBS Lett. 2004 Jun 4;567(2-3):183-8. doi: 10.1016/j.febslet.2004.04.054.

Abstract

The folding of HIV gp41 into a 6-helix bundle drives virus-cell membrane fusion. To examine the structural relationship between the 6-helix bundle core domain and other regions of gp41, we expressed in Escherichia coli, the entire ectodomain of HIV-2(ST) gp41 as a soluble, trimeric maltose-binding protein (MBP)/gp41 chimera. Limiting proteolysis indicated that the Cys-591-Cys-597 disulfide-bonded region is outside a core domain comprising two peptides, Thr-529-Trp-589 and Val-604-Ser-666. A biochemical examination of MBP/gp41 chimeras encompassing these core peptides indicated that the N-terminal polar segment, 521-528, and C-terminal membrane-proximal segment, 658-666, cooperate in stabilizing the ectodomain. A functional interaction between sequences outside the gp41 core may contribute energy to membrane fusion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Carrier Proteins / chemistry
  • Carrier Proteins / genetics
  • Chromatography, Gel
  • Cysteine / chemistry
  • Cysteine / genetics
  • Gene Expression
  • HIV Envelope Protein gp41 / biosynthesis*
  • HIV Envelope Protein gp41 / chemistry*
  • HIV Envelope Protein gp41 / genetics
  • HIV-2 / chemistry*
  • Hot Temperature
  • Humans
  • Maltose-Binding Proteins
  • Molecular Sequence Data
  • Peptide Fragments / chemistry
  • Protein Denaturation
  • Protein Structure, Quaternary
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Sequence Alignment
  • Spectrometry, Mass, Electrospray Ionization
  • Ultracentrifugation / methods

Substances

  • Carrier Proteins
  • HIV Envelope Protein gp41
  • Maltose-Binding Proteins
  • Peptide Fragments
  • Recombinant Fusion Proteins
  • Cysteine