Table 2.
Residue | Cell-Cell Fusion | Virus Entry | ||||||
---|---|---|---|---|---|---|---|---|
gp120a Expression | gp41b Expression | Fusione Category | Effectf of Alanine Mutation | gp120c Level | gp41d Level | Fusione Category | Effectf of Alanine Mutation | |
L660A | +++ | ++++ | WT | - | +++++ | +++++ | WT | - |
L661A | +++++ | +++++ | WT | - | +++++ | +++++ | WT | - |
E662A | +++++ | +++++ | WT | - | +++++ | +++++ | WT | - |
L663A | +++ | +++++ | Abolished | Severe defect | +++++ | +++++ | WT | - |
D664A | +++++ | +++++ | WT | - | ++++ | +++++ | Diminished | - |
K665A | +++ | +++++ | WT | - | +++++ | +++++ | Diminished | Mild defect |
W666A | +++++ | +++++ | Abolished | Severe defect | ++++ | ++ | Diminished | - |
S668A | +++ | +++++ | WT | - | +++++ | +++++ | WT | - |
L669A | +++++ | +++++ | WT | - | +++++ | +++++ | Enhanced | Increased Env Expression |
W670A | +++++ | +++++ | WT | - | +++++ | +++++ | Diminished | Mild defect |
N671A | + | +++ | Abolished | - | + | +++++ | Diminished | Expression/Association |
W672A | ++++ | +++++ | WT | - | ++ | ++++ | WT | Association |
F673A | +++ | +++++ | WT | +++++ | ++++ | Diminished | Mild defect | |
D674A | +++++ | +++++ | Diminished | Mild defect | ++ | ++ | WT | - |
I675A | ++++ | +++++ | WT | - | +++++ | +++++ | WT | - |
T676A | ++++ | +++++ | WT | - | +++++ | +++++ | Diminished | Mild defect |
K677A | +++++ | +++++ | WT | - | +++++ | +++++ | WT | - |
W678A | ++ | +++++ | Enhanced | Increased Env Expression | +++++ | ++++ | WT | - |
L679A | + | +++ | WT | - | +++++ | +++++ | Diminished | Mild defect |
W680A | + | +++ | Enhanced | Enhanced fusogenicity | +++++ | +++++ | WT | - |
Y681A | ++ | +++ | Enhanced | Enhanced fusogenicity | +++++ | ++++ | Diminished | Mild defect |
I682A | + | +++ | WT | - | +++++ | +++++ | Enhanced | Increased Env Expression |
K683A | ++++ | +++++ | Enhanced | Increased Env Expression | ++++ | ++++ | Diminished | Mild defect |
Protein (gp120 a, and gp41 b) levels were shown in the cell lysates used for cell–cell fusion and
Protein (gp120 c, and gp41 d) levels of expression shown in virus. The levels of protein expression were shown as follows: +, 0–20% of WT levels; ++, 20–50% of WT levels; +++, 50–80% of WT levels; ++++, >80% of WT levels; and +++++, >120% of WT levels.
Fusion categories were defined as follows: WT (no effect, WT level), diminished (20–80% of wild type), abolished (<20% of wild type), and enhanced ( >120% wild type).
Effect of alanine mutations were defined as follows, expression/folding (<20% of WT gp41), cleavage (<20% gp120 and gp41, WT level of gp160), association (<0.5of gp120/gp41 ratio), severe fusogenicity (>20 % of WT gp41, >0.5 gp120/gp41 ratio, < 20% of WT fusion level), mild fusogenicity (>20 % of WT gp41, >0.5 gp120/gp41 ratio, 20–80% of WT level), increased Env expression ( >120% of WT gp160 proteins and fusion level), and enhanced fusogenicity (>120% of WT fusion, WT gp160 expression level.)