Extended Data Table 3. Expression and purification conditions.
Protein | Expression | Buffer A | Buffer B |
---|---|---|---|
Bs ClpC | 37°C, 3h | 25 mM Tris pH 7.5, 300 mM NaCl | 25 mM Tris pH 7.5, 300 mM NaCl |
Bs ClpCDWB | 37°C, 3h | 25 mM Tris pH 8, 300 mM NaCl | 25 mM Tris pH 7.5, 300 mM NaCl |
Bs ClpCNTD | 37°C, 2h | 25 mM Tris pH 8.5, 150 mM NaCl, 1 mM TCEP | 5 mM Tris pH 8.5, 50 mM NaCl, 1 mM TCEP |
Ec ClpANTD | 37°C, 2h | 25 mM Tris pH 8.5, 150 mM NaCl, 1 mM TCEP | 5 mM Tris pH 8.5, 50 mM NaCl, 1 mM TCEP |
Bs ClpP | 37°C, 3h | 25 mM Tris pH 7.5, 150 mM NaCl, 10% glycerol | 25 mM Tris pH 7.5, 150 mM NaCl, 10% glycerol |
Bs McsA | 20°C, 5h | 25 mM Tris pH 7.5, 300 mM NaCl, TCEP 1 mM | 25 mM Tris pH 7.5, 100 mM NaCl, TCEP 1 mM |
Bs McsB | 18°C, O/N | 25 mM Tris pH 7.5, 300 mM KCl | 25 mM Tris pH 7.5, 50 mM KCl |
Bs McsBE212A | 18°C, O/N | 25 mM Tris pH 7.5, 300 mM KCl | 25 mM Tris pH 7.5, 50 mM KCl |
Gs McsB | 37°C, 3h | 25 mM Hepes pH 7.5, 300 mM KCl | 25 mM Hepes pH 7.5, 100 mM KCl |
Gs McsBE122A | 18°C, O/N | 50 mM Tris pH 7.5, 50 mM NaCl | 50 mM Tris pH 7.5, 50 mM NaCl |
Bs MecA | 25°C, 5h | 25 mM Hepes pH 8, 300 mM KCl | 25 mM Hepes pH 8, 300 mM KCl |
Gs YwlE | 37°C, 3h | 25 mM Tris pH 7.5, 300 mM NaCl | 25 mM Tris pH 7.5, 100 mM NaCl |
Gs YwlED118N | 37°C, 3h | 25 mM Tris pH 7.5, 300 mM NaCl | 25 mM Tris pH 7.5, 100 mM NaCl |
Bs: Bacillus subtilis, Ec: Escherichia coli, Gs: Geobacillus stearothermophilus, O/N: overnight