On the interaction between gp41 and membranes: the immunodominant loop stabilizes gp41 helical hairpin conformation.
gp41 is the protein responsible for the process of membrane fusion that allows primate lentiviruses (HIV and SIV) to enter into their host cells. gp41 ectodomain contains an N-terminal and a C-terminal heptad repeat region (NHR and CHR) connected by an immunodominant loop. In the absence of membranes, the NHR and CHR segments fold into a protease-resistant core with a trimeric helical hairpin structure. However, when the immunodominant loop is not present (either in a complex formed by HIV-1 gp41-derived NHR and CHR peptides or by mild treatment with protease of recombinant constructs of HIV-1 gp41 ectodomain, which also lack the N-terminal fusion peptide and the C-terminal Trp-rich region) membrane binding induces a conformational change in the gp41 core structure. Here, we further investigated whether covalently linking the NHR and CHR segments by the immunodominant loop affects this conformational change. Specifically, we analyzed a construct corresponding to a fragment of SIVmac239 gp41ectodomain (residues 27-149, named e-gp41) by means of surface plasmon resonance, Trp and rhodamine fluorescence, ATR-FTIR spectroscopy, and differential scanning calorimetry. Our results suggest that the presence of the loop stabilizes the trimeric helical hairpin both when e-gp41 is in aqueous solution and when it is bound to the membrane surface. Bearing in mind possible differences between HIV-1 and SIV gp41, and considering that the gp41 ectodomain constructs analyzed to date lack the N-terminal fusion peptide and the C-terminal Trp-rich region, we discuss our observations in relation to the mechanism of virus-induced membrane fusion.
Peisajovich
Sergio G
SG
Department of Biological Chemistry, Weizmann Institute of Science, 76100, Rehovot, Israel.
Blank
Lior
L
Epand
Raquel F
RF
Epand
Richard M
RM
Shai
Yechiel
Y
D002152:Calorimetry, Differential Scanning; D002462:Cell Membrane / Q000502:physiology*; D002942:Circular Dichroism; D003545:Cysteine / Q000737:chemistry; D005453:Fluorescence; D006358:Hot Temperature; D006801:Humans; D016056:Immunodominant Epitopes / Q000737:chemistry* / Q000378:metabolism; D008081:Liposomes; D008561:Membrane Fusion; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000378:metabolism; D008954:Models, Biological; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D012191:Retroviridae Proteins / Q000737:chemistry* / Q000378:metabolism; D012235:Rhodamines / Q000737:chemistry / Q000378:metabolism; D015302:Simian Immunodeficiency Virus / Q000378:metabolism*; D017550:Spectroscopy, Fourier Transform Infrared; D020349:Surface Plasmon Resonance; D014364:Tryptophan / Q000737:chemistry*
D016056:Immunodominant Epitopes; D008081:Liposomes; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D012191:Retroviridae Proteins; D012235:Rhodamines; C082614:SIV envelope protein gp41; D014364:Tryptophan; D003545:Cysteine