TABLE 1.
Effects of AD101 and SCH-C on HIV-1 entry mediated by CCR5 extracellular domain mutantsa
| Domain | Mutant | Relative % entry in presence of:
|
|
Domain | Mutant | Relative % entry in presence of:
|
||
|---|---|---|---|---|---|---|---|---|
| AD101 | SCH-C | AD101 | SCH-C | |||||
| Nt | D2 | 0.0 ± 0.0 | 0.0 ± 0.0 | ECL2 | R168 | 0.3 ± 0.8 | 0.0 ± 0.0 | |
| Y3 | 0.0 ± 0.0 | 0.0 ± 0.0 | S169 | NA | NA | |||
| Q4 | 0.0 ± 0.0 | 0.0 ± 0.0 | Q170 | 0.5 ± 0.8 | 2.5 ± 0.6 | |||
| S6 | 0.3 ± 2.7 | 0.0 ± 0.0 | K171A/E172A | 0.0 ± 0.0 | 0.0 ± 0.0 | |||
| S7 | 0.3 ± 4.2 | 0.2 ± 1.4 | H175 | 0.0 ± 0.0 | 0.0 ± 0.0 | |||
| P8 | 0.8 ± 0.4 | 1.3 ± 2.1 | Y176 | 0.0 ± 0.0 | 0.0 ± 0.0 | |||
| Y10 | 0.0 ± 0.0 | 0.0 ± 0.0 | T177 | 0.0 ± 0.0 | 0.0 ± 0.0 | |||
| D11 | 0.0 ± 0.0 | 0.0 ± 0.0 | C178 | 2.7 ± 2.3 | 2.6 ± 2.2 | |||
| N13 | 0.0 ± 0.0 | 0.0 ± 0.0 | S179 | 1.5 ± 1.8 | 0.0 ± 0.0 | |||
| Y14 | 0.4 ± 1.5 | 0.0 ± 0.0 | S180 | 0.7 ± 2.3 | 1.8 ± 0.8 | |||
| Y15 | NA | NA | H181 | 0.0 ± 0.0 | 0.0 ± 0.0 | |||
| T16 | 0.0 ± 0.0 | 0.0 ± 0.0 | F182 | 4.0 ± 1.0 | 3.4 ± 0.9 | |||
| S17 | 0.0 ± 0.0 | 0.0 ± 0.0 | P183 | 1.0 ± 1.9 | 4.2 ± 1.1 | |||
| E18 | 0.0 ± 0.0 | 0.0 ± 0.0 | Y184 | 0.0 ± 0.0 | 0.0 ± 0.0 | |||
| P19 | 0.0 ± 0.0 | 0.0 ± 0.0 | S185 | 0.0 ± 0.0 | 0.0 ± 0.0 | |||
| C20 | 0.9 ± 1.0 | 0.0 ± 0.0 | Q186 | 0.0 ± 0.0 | 0.0 ± 0.0 | |||
| Q21 | 0.0 ± 0.0 | 0.0 ± 0.0 | Y187 | 1.2 ± 0.1 | 0.0 ± 0.0 | |||
| K22 | 0.0 ± 0.0 | 0.0 ± 0.0 | Q188 | 0.0 ± 0.0 | 0.0 ± 0.0 | |||
| N24 | 0.0 ± 0.0 | 0.0 ± 0.0 | F189 | 0.0 ± 0.0 | 0.0 ± 0.0 | |||
| K26 | 0.0 ± 0.0 | 0.2 ± 1.5 | W190 | 0.0 ± 0.0 | 0.0 ± 0.0 | |||
| Q27 | 0.0 ± 0.0 | 0.0 ± 0.0 | K191A/N192A | 0.9 ± 1.3 | 3.8 ± 0.8 | |||
| R31 | 2.9 ± 1.2 | 0.0 ± 0.0 | F193 | 0.1 ± 0.7 | 0.0 ± 0.0 | |||
| ECL1 | H88 | 0.4 ± 1.6 | 0.9 ± 0.2 | Q194 | 0.3 ± 0.5 | 0.0 ± 0.0 | ||
| Y89 | 0.0 ± 0.0 | 0.0 ± 0.0 | ECL3 | E262 | 0.0 ± 0.0 | 0.0 ± 0.0 | ||
| Q93 | 1.7 ± 1.5 | 1.5 ± 0.2 | F263 | 0.0 ± 0.0 | 0.0 ± 0.0 | |||
| W94 | 0.0 ± 0.0 | 0.0 ± 0.0 | F264 | 0.0 ± 0.0 | 0.0 ± 0.0 | |||
| D95 | 0.0 ± 0.0 | 0.0 ± 0.0 | N267 | 0.0 ± 0.0 | 0.0 ± 0.0 | |||
| F96 | 0.0 ± 0.0 | 4.2 ± 2.8 | N268 | 0.0 ± 1.8 | 0.0 ± 0.0 | |||
| N98 | 1.3 ± 1.6 | 3.7 ± 1.4 | C269 | 0.0 ± 0.0 | 0.0 ± 0.0 | |||
| T99 | 0.2 ± 2.2 | 0.0 ± 0.0 | S270 | 0.1 ± 2.8 | 0.0 ± 0.0 | |||
| M100 | 0.5 ± 1.7 | 0.0 ± 0.0 | S271 | 0.0 ± 0.0 | 0.0 ± 0.0 | |||
| C101 | NA | NA | S272 | 0.0 ± 0.0 | 0.0 ± 0.0 | |||
| Q102 | 0.0 ± 0.0 | 0.0 ± 0.0 | N273 | 0.0 ± 0.0 | 0.0 ± 0.0 | |||
| R274A/D276A | 2.2 ± 1.6 | 6.3 ± 1.0 | ||||||
| Q277 | 0.6 ± 2.1 | 0.0 ± 0.0 | ||||||
a
Entry of HIV-1JR-FL Env-pseudotyped reporter viruses into U87-CD4 cells transiently expressing CCR5 alanine mutants was measured in the presence of 100 nM AD101 or SCH-C. Relative HIV-1 entry in the presence of inhibitors was calculated as (RLU with inhibitor/RLU without inhibitor) × 100%. All values are means ± SD of three independent experiments. NA, not applicable; alanine substitutions of residues Y15 in the Nt, C101 in ECL1, and S169 in ECL2 were excluded from the study because they are substantially defective at mediating HIV-1 entry (<1% of wild-type CCR5 coreceptor function) (12, 13).