TABLE 5.
Site-directed mutants of NleB1 and their effect on FADD GlcNAcylation and binding
| Mutant | Ability to GlcNAcylate FADDa | Binding to FADDa |
|---|---|---|
| NleB1 single-site-directed mutants | ||
| P63A | +++ | +++ |
| I64A | +++ | +++ |
| L65A | +++ | +++ |
| N66A | +++ | +++ |
| K68A | +++ | +++ |
| K81A | +++ | +++ |
| D93A | +++ | + |
| D121A | +++ | +++ |
| D128A | +++ | − |
| L157A | +++ | + |
| Y219A | − | +++ |
| Y234A | +++ | +++ |
| P236A | +++ | +++ |
| D237A | +++ | +++ |
| G238A | +++ | + |
| I239A | +++ | + |
| H242A | +++ | +++ |
| E253A | − | +++ |
| N263A | +++ | +++ |
| L267A | +++ | + |
| A269E | +++ | +++ |
| Y283A | +++ | + |
| K292A | +++ | + |
| NleB1 multiple-site-directed mutants | ||
| PILN(63–66)AAAA | − | +++ |
| HKQ(140–142)AAA | +++ | +++ |
| QES(142–144)AAA | +++ | ++ |
| LGLL(155–158)AAAA | +++ | − |
| GSL(197–199)AAA | +++ | + |
| DKL(228–230)AAA | +++ | − |
| PDG(236–238)AAA | + | − |
| SN(262–263)AA | +++ | ++ |
| LAGL(268–271)AEAA | +++ | − |
| KV(277–278)AA | +++ | +++ |
| DAD(221–223)AAA | − | +++ |
| KGI(289–291)AAA | +++ | − |
a
+, ++, and +++, increasing levels of FADD GlcNAcylation or binding, with +++ reflecting the level of FADD GlcNAcylation or binding observed in the presence of wild-type NleB1; −, an absence of FADD GlcNAcylation or binding