Table 1.
List of NMR samples used in the present study
| Sample | Labeling | Buffer | Construct | NMR experiment |
|---|---|---|---|---|
| A1–A5a | AILMV | 99.99% 2H2O, 100 mM NaCl, 0.1× EDTA-free complete protease inhibitor (Roche), and 20 mM Tris-pH 7.4 | ~0.2 mM EIN-R1p | PRE experiment |
| B | AILV | 99.99% 2H2O, 100 mM NaCl, 0.1× EDTA-free complete protease inhibitor (Roche), and 20 mM Tris-pH 7.4 | ~1 mM WT EIN | 1H–13C HMQC-NOESY-HMQC |
| C | Uniformly 15N + ILV (linearized spin systems) | 5% 2H2O/95% H2O, 100 mM NaCl, 0.1× EDTA-free complete protease inhibitor (Roche), and 20 mM Tris-pH 7.4 | ~1 mM WT EIN | HC(CO)NH-TOCSY |
| D | A | 99.99% 2H2O, 100 mM NaCl, 0.1× EDTA-free complete protease inhibitor (Roche), and 20 mM Tris-pH 7.4 | ~0.2 mM WT EIN | 1H–13C HMQC |
| E | I | 99.99% 2H2O, 100 mM NaCl, 0.1× EDTA-free complete protease inhibitor (Roche), and 20 mM Tris-pH 7.4 | ~0.2 mM WT EIN | 1H–13C HMQC |
| F | M | 99.99% 2H2O, 100 mM NaCl, 0.1× EDTA-free complete protease inhibitor (Roche), and 20 mM Tris-pH 7.4 | ~0.2 mM WT EIN | 1H–13C HMQC |
| G | LV | 99.99% 2H2O, 100 mM NaCl, 0.1× EDTA-free complete protease inhibitor (Roche), and 20 mM Tris-pH 7.4 | ~0.2 mM WT EIN | 1H–13C HMQC |
All samples are in a uniform 2H, 12C and 14N background unless otherwise specified. Methyl-bearing side chains with 1H–13C labeled methyl groups are listed in the second column
a
The five samples correspond to R1p conjugated to surface engineered cysteines at A52C, Q87C, S113C, S196C and A241C