Methods used to examine variant α globin interactions
| Interaction/method . | WT . | R31S . | K99E . | K99N . | H103Y . | H103R . | F117S . | P119S . | A130D . |
|---|---|---|---|---|---|---|---|---|---|
| AHSP-α | |||||||||
| Coexpression in E coli | ++ | ++ | + | + | + | − | + | − | ++ |
| GST pulldown | ++ | + | − | − | − | − | − | − | ++ |
| Limited proteolysis | ++ | − | − | − | − | − | − | − | + |
| α1β1 | |||||||||
| Monomer shift assay | ++ | − | ++ | ++ | + | − | ND | ++ | ++ |
| Coexpression in E coli | ++ | − | ++ | ++ | − | − | − | − | ++ |
| Summary of biochemical effects | |||||||||
| Binding to AHSP | ++ | ++ | + | + | + | − | + | − | ++ |
| Binding to β globin | ++ | − | ++ | ++ | + | − | + | + | ++ |
| Interaction/method . | WT . | R31S . | K99E . | K99N . | H103Y . | H103R . | F117S . | P119S . | A130D . |
|---|---|---|---|---|---|---|---|---|---|
| AHSP-α | |||||||||
| Coexpression in E coli | ++ | ++ | + | + | + | − | + | − | ++ |
| GST pulldown | ++ | + | − | − | − | − | − | − | ++ |
| Limited proteolysis | ++ | − | − | − | − | − | − | − | + |
| α1β1 | |||||||||
| Monomer shift assay | ++ | − | ++ | ++ | + | − | ND | ++ | ++ |
| Coexpression in E coli | ++ | − | ++ | ++ | − | − | − | − | ++ |
| Summary of biochemical effects | |||||||||
| Binding to AHSP | ++ | ++ | + | + | + | − | + | − | ++ |
| Binding to β globin | ++ | − | ++ | ++ | + | − | + | + | ++ |
++ indicates normal interaction; +, reduced interaction; −, no interaction detected; and ND, not determined.