Table 6.
Nomenclature and description of HIV-1 gp160 constructs
All gp160 constructs were expressed in mammalian cells.
| No. | Construct name | Description |
|---|---|---|
| 1a | WT gp160 | Full-length WT gp160 from the JRFL strain with all PNGS intact. It contains 15 PNGS in the core gp120 region |
| 2 | ΔG11 gp160 | A WT gp160 variant lacking 11 OD PNGS. It contains only four core gp120 PNGS: two at the interface of the inner and outer domains of gp120 (at residues 262 and 276) and two in the inner domain of gp120 (at residues 88 and 241) |
| 3 | ΔG12 gp160 | Asn-276 glycan removed in ΔG11 gp160 background. |
| 4 | ΔG13 gp160 | Devoid of all OD PNGS including two (Asn-262 and Asn-276) present at the interface of inner and outer domains of gp120 |
| 5 | ΔG14 gp160 | Devoid of all OD PNGS + mutation at inner domain PNGS position 88 (N88G) |
| 6 | ΔG15F gp160 | Devoid of all 15 core gp120 glycans. Mutations at inner domain positions 88 (N88G) and 241 (N241H) in this construct were selected using yeast library screening of core gp120 |
| 7 | T332N ΔG15F gp160 | ΔG15F gp160 with T332N mutation to reintroduce PGT128 bNAb epitope |
| 8 | ΔGQ11 gp160 | A variant of ΔG11 gp160 wherein Asn residues at 11 OD PNGS were mutated to structurally similar Gln instead of rational mutations |
| 9 | SEKS gp160 | Cleavage defective JRFL gp160. Furin cleavage site REKR is mutated to SEKS in WT gp160 background |
| 10b | Q842 WT gp160 | WT gp160 from subtype A strain Q842env.d16 |
| 11 | Q842 ΔG13 gp160 | Gln-842 gp160 variant lacking 13 OD PNGS |
| 12 | QH343 WT gp160 | WT gp160 from subtype A strain QH343.21M.ENV.B5 |
| 13 | QH343 ΔG12 gp160 | QH343 gp160 variant lacking 12 OD PNGS |
| 14 | DU WT gp160 | WT gp160 from subtype A strain DU422.1 |
| 15 | DU ΔG10 gp160 | DU gp160 variant lacking 10 OD PNGS |
| 16 | CAP WT gp160 | WT gp160 from subtype A strain CAP45.2.00G3 |
| 17 | CAP ΔG12 gp160 | CAP gp160 variant lacking 12 OD PNGS |
a Constructs 1–9 are based on the JRFL isolate. All JRFL gp160 molecules have the E168K mutation. The E168K mutation confers PG9/PG16 (trimer-specific bNAbs) binding on JRFL Env. All JRFL gp160 constructs have a cytoplasmic tail deletion (ΔCT) that is known to improve expression on the mammalian surface. Mutations introduced in JRFL gp160 constructs were the same as in core gp120 constructs and are described in supplemental Table S3.
b Constructs 10–17 are based on different HIV-1 strains from clade A and clade C HIV-1 reference panel of Env clones. These construct names bear the initial of the strain from which they are derived. A variable number of OD glycosylation sites are present in different strains, and the nomenclature reflects the number of PNGS mutations in each construct. Mutations introduced in these gp160 variants are described in supplemental Table S4.