Design and properties of N(CCG)-gp41, a chimeric gp41 molecule with nanomolar HIV fusion inhibitory activity

J Biol Chem. 2001 Aug 3;276(31):29485-9. doi: 10.1074/jbc.C100317200. Epub 2001 Jun 19.


The design and characterization of a chimeric protein, termed N(CCG)-gp41, derived from the ectodomain of human immunodeficiency virus (HIV), type I gp41 is described. N(CCG)-gp41 features an exposed trimeric coiled-coil comprising the N-terminal helices of the gp41 ectodomain. The trimeric coiled-coil is stabilized both by fusion to a minimal thermostable ectodomain of gp41 and by engineered intersubunit disulfide bonds. N(CCG)-gp41 is shown to inhibit HIV envelope-mediated cell fusion at nanomolar concentrations with an IC(50) of 16.1 +/- 2.8 nm. It is proposed that N(CCG)-gp41 targets the exposed C-terminal region of the gp41 ectodomain in its pre-hairpin intermediate state, thereby preventing the formation of the fusogenic form of the gp41 ectodomain, which comprises a highly stable trimer of hairpins arranged in a six-helix bundle. N(CCG)-gp41 has potential as a therapeutic agent for the direct inhibition of HIV cell entry, as an anti-HIV vaccine, and as a component of a rapid throughput assay for screening for small molecule inhibitors of HIV envelope-mediated cell fusion. It is anticipated that antibodies raised against N(CCG)-gp41 may target the trimeric coiled-coil of N-terminal helices of the gp41 ectodomain that is exposed in the pre-hairpin intermediate state in a manner analogous to peptides derived from the C-terminal helix of gp41 that are currently in clinical trials.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 3T3 Cells
  • AIDS Vaccines / biosynthesis
  • AIDS Vaccines / chemistry
  • Amino Acid Sequence
  • Animals
  • Anti-HIV Agents / chemistry*
  • Anti-HIV Agents / pharmacology
  • Base Sequence
  • CD4 Antigens / physiology
  • Cell Fusion*
  • Cell Line
  • Circular Dichroism
  • Crystallography, X-Ray
  • Disulfides
  • Drug Design
  • HIV Envelope Protein gp41 / biosynthesis*
  • HIV Envelope Protein gp41 / chemistry
  • HIV Envelope Protein gp41 / genetics*
  • HIV-1 / drug effects
  • HIV-1 / genetics
  • HIV-1 / physiology*
  • Humans
  • Mice
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Conformation
  • Protein Engineering
  • Protein Folding
  • Protein Structure, Secondary
  • Recombinant Fusion Proteins / biosynthesis*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / pharmacology
  • Spectrophotometry, Ultraviolet
  • Viral Envelope Proteins / physiology


  • AIDS Vaccines
  • Anti-HIV Agents
  • CD4 Antigens
  • Disulfides
  • HIV Envelope Protein gp41
  • Recombinant Fusion Proteins
  • Viral Envelope Proteins