Conserved arginine residue in the membrane-spanning domain of HIV-1 gp41 is required for efficient membrane fusion.
10.1007/s13238-011-1051-0
- Author:
Yufei LONG
1
;
Fanxia MENG
;
Naoyuki KONDO
;
Aikichi IWAMOTO
;
Zene MATSUDA
Author Information
1. China-Japan Joint Laboratory of Structural Virology and Immunology, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.
- Publication Type:Journal Article
- MeSH:
Amino Acid Substitution;
Animals;
Arginine;
chemistry;
metabolism;
Cell Line;
Cercopithecus aethiops;
HIV Envelope Protein gp41;
chemistry;
metabolism;
HIV-1;
metabolism;
Humans;
Kinetics;
Membrane Fusion;
physiology;
Mutation;
Protein Structure, Tertiary
- From:
Protein & Cell
2011;2(5):369-376
- CountryChina
- Language:English
-
Abstract:
Despite the high mutation rate of HIV-1, the amino acid sequences of the membrane-spanning domain (MSD) of HIV-1 gp41 are well conserved. Arginine residues are rarely found in single membrane-spanning domains, yet an arginine residue, R(696) (the numbering is based on that of HXB2), is highly conserved in HIV-1 gp41. To examine the role of R(696), it was mutated to K, A, I, L, D, E, N, and Q. Most of these substitutions did not affect the expression, processing or surface distribution of the envelope protein (Env). However, a syncytia formation assay showed that the substitution of R(696) with amino acid residues other than K, a naturally observed mutation in the gp41 MSD, decreased fusion activity. Substitution with hydrophobic amino acid residues (A, I, and L) resulted in a modest decrease, while substitution with D or E, potentially negatively-charged residues, almost abolished the syncytia formation. All the fusion-defective mutants showed slower kinetics with the cell-based dual split protein (DSP) assay that scores the degree of membrane fusion based on pore formation between fusing cells. Interestingly, the D and E substitutions did show some fusion activity in the DSP assays, suggesting that proteins containing D or E substitutions retained some fusion pore-forming capability. However, nascent pores failed to develop, due probably to impaired activity in the pore enlargement process. Our data show the importance of this conserved arginine residue for efficient membrane fusion.
