TABLE 2.
Effects of radical LcrV mutations on the secretion of Yopsa
| LcrV | Secretion in medium:
|
|
|---|---|---|
| Without Ca2+ | With Ca2+ | |
| None | Blocked | Blocked |
| Wild type | Activated | Blocked |
| L291R | Activated | Blocked |
| I294R | Activated | Blocked |
| F298R | Activated | Blocked |
| A301R | Activated | Blocked |
| I302R | Activated | Blocked |
| A304R | Activated | Blocked |
| L305R | Activated | Blocked |
| F308Rb | Reduced | Blocked |
| I309R | Activated | Blocked |
| L291R I294R | Activated | Blocked |
| L291R F298R | Activated | Blocked |
| L291R A301R | Activated | Blocked |
| L291R I302R | Activated | Blocked |
| I294R F298R | Activated | Blocked |
| A301R L305R | Activated | Blocked |
| I302R L305R | Activated | Blocked |
| L291R F308Rc | Blocked | Blocked |
| I294R F308Rb | Reduced | Blocked |
| A301R F308Rb | Reduced | Blocked |
| A301R F308Rb | Reduced | Blocked |
| I302R F308Rb | Reduced | Blocked |
| L305R F308Rb | Reduced | Blocked |
a
Secretion phenotype of Yops produced by the ΔlcrV5 strain expressing various LcrV radical mutants. ΔlcrV5 strains expressing various LcrV constructs were grown in TMH for 4 h at 37°C in the presence or absence, of Ca2+ and the cell-free fractions were analyzed by SDS-PAGE followed by silver staining.
b
Mutation resulting in an LcrV mutant that showed reduced Yop secretion.
c
Mutation resulting in an LcrV mutant that showed no Yop secretion.