Thermodynamics of trimer-of-hairpins formation by the SIV gp41 envelope protein

Ilian Jelesarov, Min Lu

Research output: Contribution to journalArticlepeer-review

40 Scopus citations

Abstract

The gp41 envelope protein mediates the entry of primate immunodeficiency viruses into target cells by promoting the fusion of viral and cellular membranes. The structure of the gp41 ectodomain core represents a trimer of identical helical hairpins in which a central trimeric coiled-coil made up of three amino-terminal helices is wrapped in an outer layer of three antiparallel carboxyl-terminal helices. Triggering formation of this fusion-active gp41 conformation appears to cause close membrane apposition and thus overcome the activation energy barrier for lipid bilayer fusion. We present a detailed description of the folding thermodynamics of the simian immunodeficiency virus (SIV) gp41 core by using a recombinant trimeric N34(L6)C28 model. Differential scanning calorimetry and spectroscopic experiments on denaturant-induced and thermal unfolding indicate that the free energy of association of three 68 residue N34(L6)C28 peptides to a trimer-of-hairpins is 76 kJ mol-1 at pH 7.0 and 25 °C in physiological buffer. The associated enthalpy change, ΔHunf, is 177 kJ mol-1, while the entropy of unfolding, ΔSunf, is 0.32 kJ K-1 mol-1 The temperature of maximal stability is close to 20°C. The unfolding heat capacity increment is ∼9 kJ K-1 mol-1 (∼45 J K-1 mol residue-1), which is lower than expected for unfolding of the trimer to an extended and fully hydrated polypeptide chain. Replacement by isoleucine of the polar residues Thr582 or Thr586 buried in the N-terminal trimeric coiled-coil interface leads to very strong stabilization of the trimer-of-hairpins, 30-35 kJ mol-1. Single-point mutations in the central coiled-coil thus strongly stabilize the gp41 core structure. These thermodynamic characteristics may be important for the refolding of the gp41 envelope protein into its fusion-active conformation during membrane fusion.

Original languageEnglish (US)
Pages (from-to)637-656
Number of pages20
JournalJournal of molecular biology
Volume307
Issue number2
DOIs
StatePublished - Mar 23 2001

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Molecular Biology

Keywords

  • Coiled-coil
  • Envelope glycoprotein
  • Gp41
  • Membrane fusion
  • Trimer-of-hairpins

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