Table 2. Summary of oligomeric states, enzymatic activity, ESCRT-III disassembly and HIV-1 budding of MsVps4 and hVPS4A, and B constructs.
| MsVps4 and hVPS4 constructs | Domain | Elution volumes (ml) | Complex* | ATPase† | ESCRT-III‡ | HIV-1§ | |||
|---|---|---|---|---|---|---|---|---|---|
| MsVps4_wild-type | 11.6 | 13.4 | 15.8 | 17.1 | Dodecamer | 100% | — | — | |
| hVPS4B_E235Q +ATP | 14.1 | 17.8 | Hexamer | — | — | — | |||
| hVPS4B_wild-type +ATP/Mg | — | — | — | — | — | 100% | 100% | ||
| hVPS4A_wild type | — | — | — | — | — | — | — | 100% | |
| MsVps4_Phe126Ala | H | 16.4 | Monomer | 0.0±0.1% | — | — | |||
| hVPS4B_E235Q_Phe160Ala+ATP | H | 14.7 | 16.7 | 17.9 | Monomer | — | — | — | |
| hVPS4B_Phe160Ala+ATP/Mg | H | — | — | — | — | — | 0.0±0.1% | Inhibition | — |
| hVPS4A_Phe153Ala | H | — | — | — | — | — | — | — | Inhibition |
| MsVps4_Met318Ala | H | 13.1 | 14.6 | 17.2 | Hexamer | 154±2,5% | — | — | |
| hVPS4B_E235Q_Met350Ala+ATP | H | 14.3 | 16.6 | Monomer | — | — | |||
| hVPS4B_Met350Ala+ATP/Mg | H | — | — | — | — | — | 0.0±0.1% | ∼20% | — |
| MsVps4_Ile89Glu | AAA | 12.6 | 15.8 | 17.5 | Dodecamer | 85.6±16.4% | — | — | |
| hVPS4B_E235Q_Ile124Glu+ATP | AAA | 13.6 | 18 | Hexamer | — | — | — | ||
| hVPS4B_Ile124Glu+ATP/Mg | AAA | — | — | — | — | — | 8.10±1.95% | Inhibition | — |
| hVPS4A_Val117Glu | AAA | — | — | — | — | — | — | — | 100% |
| MsVps4_Glu174Ala_Glu176Ala | V | 10.5 | 13.7 | 16 | 17.6 | Dodecamer | 0.14±0.1% | — | — |
| hVPS4A_Glu204Ala_Glu206Ala | V | — | — | — | — | — | — | — | Inhibition |
| MsVps4_Arg259Ala_Arg260Ala | V | 13.7 | 15.4 | 17.3 | Hexamer | 0.1±0.02% | — | ||
| MsVps4_Glu214Tyr | pore loop 2 | 10.7 | 13.5 | 15.9 | 17.6 | Dodecamer | 7.9±1.5% | — | — |
| hVPS4A_Glu240Tyr | pore loop 2 | — | — | — | — | — | — | — | Inhibition |
| MsVps4_Ser169Ala | V | 11.9 | 14.6 | — | 17.4 | Dodecamer | 14.5±3.2% | — | — |
| hVPS4B_Arg253Ala+ATP/Mg | V | — | — | — | — | — | 69.5±5% | ∼50% | — |
*Main complex formed in vitro.
†ATPase activity; reported ATPase values correspond to the mean±s.d. of at least three distinct experiments carried out on two or more batches of MsVps4.
‡Disassembly of ESCRT-III CHMP2A-CHMP3 tubular structures.
§Inhibition of HIV-1 budding.