A cross-neutralizing antibody between HIV-1 and influenza virus
Fig 5
Glycoforms at N165 and N246 and 2G12 binding mode to HA wildtype and glycan mutants.
(A) Site-specific glycosylation analysis of Bris07 HA glycan knockout mutants at N165 and N246 (left panel). Percentage differences in abundance of oligomannose-type glycans between glycan knockout mutants and WT Bris07 HA is also illustrated with decreased and increased abundance colored in red and blue (right panel), respectively. (B) Negative-stain electron microscopy (nsEM) images show the different modes of 2G12 recognition of Bris07 wild type (left panel) and N165A mutant (right panel) HA. Fab 2G12, as a domain-swapped dimer, and the HA trimer are indicated by arrows. 2G12 Fab involved in binding to two N246 glycans, one N246 glycan, and one N165 and possibly one N246 glycan are shown in green, purple, and yellow, respectively. Wild type and N165A HA are in white. (C) Front view of A/Brisbane/10/07 (H3N2) HA trimer (PDB 6AOV, in grey) [91]. Man9GlcNAc2 on N165 and N246 were modelled onto Bris07 wild type HA with Charmm-Gui [92]. 2G12 Fab dimer (PDB 6N2X, in cyan) [11] is shown bound to the lateral side (left panel) and apex of trimer in parallel (middle panel) or tilted (right panel) binding mode from the nsEM data in Fig 5B. Man9GlcNAc2 on N165 and N246 are in yellow and green, respectively. (D) Same as (C), but N165A is shown in magenta. N246 was modelled with Man8GlcNAc2. 2G12 is shown bound to the apex of Bris07 N165A trimer as in the nsEM reconstruction. The Man8GlcNAc2 at N246 is in green.