Table 2. Convertibility of the PrP glycosylation mutants in cell culture and by cell PMCA.
| Glycotype | Genotype | Cell culture conversion* | Cell-PMCA conversion | Incubation time of tg338 mice inoculated with Cell-PMCA amplicons in days ± SEM (n/n0) |
|---|---|---|---|---|
| Wild type (VRQ) | Wild type | + | + | 70 ± 1 (5/5) |
| Monoglycosylated (Site 1) | N184D | + | + | 68 ± 1 (5/5) |
| N184Q | _ | + | 77 ± 1 (5/5) | |
| Monoglycosylated (Site 2) | N200D | + | + | 69 ± 1 (5/5) |
| N200Q | + | + | nd | |
| Unglysosylated | N184D-N200D | − | + | 68 ± 1 (5/5) |
| Unglysosylated mutants → tg338 brain PMCA# | + | 65 ± 1 (5/5) | ||
| Unseeded Wild type (VRQ) | none | − | >250 (0/5) |
n/n0: number of mice with neurological disease and positive for PrPres in the brain by immunoblotting/number of inoculated tg338 mice. The seeds used to infect mice were from a 127S 10−7 seed amplified over 2 rounds, so as to avoid any residual input.
*Cell conversion was assessed in ref. 14.
#The PMCA product inoculated was obtained by seeding tg338 brain lysate with unglycosylated PrPSc seed (10−7 dilution), itself obtained by seeding the NDND double mutant cell lysate with 10−8 diluted 127S seed. nd: not done.