Table 1.
Data collection, phasing and refinement statistics
| WT-SeMet | H171A–ADS | H171N–AMP•FUM | |
|---|---|---|---|
| A. Data collection | |||
| Wavelength (λ) | 0.9794 | 1.5418 | 1.5418 |
| f′, f″ | −8.30, 3.55 | — | — |
| Space group | I222 | P21212 | P21212 |
| Unit cell dimensions (Å) | a=68.4, b=98.4, c=136.2 | a=98.4, b=143.2, c=69.5 | a=99.1 b=143.4, c=69.5 |
| Monomers per ASU | 1 | 2 | 2 |
| Resolution limits (Å) | 50.00–2.00 | 44.00–1.85 | 35–2.0 |
| Total data | 637,617 | 782,190 | 579,421 |
| Unique data | 60,017 | 84,199 | 67,697 |
| Mean redundancy | 10.6 (10.5)b | 9.29 (6.08)b | 8.56 (8.38)b |
| Completeness (%) | 100.0 (100.0)b | 99.7 (97.0)b | 99.9 (100.0)b |
| Average I/σI | 88.9 (58.8)b | 19.5 (5.6)b | 19.9 (7.4)b |
| % reflections for which I>3σI | 100.0 (100.0)b | 98.8 (95.2)b | 98.5 (97.8)b |
| Rsym (%)a | 4.8 (7.6)b | 6.8 (30.8)b | 6.7 (25.7)b |
| B. Phasing | |||
| Phasing power | 3.18 | — | — |
| FOM before solvent flattening | 0.512 | — | — |
| FOM after solvent flattening | 0.941 | — | — |
| C. Refinement | |||
| Resolution range (Å) | 50.00–2.00 | 44.00–1.85 | 35–2.0 |
| Rcryst (%)c | 16.5 | 18.3 | 17.5 |
| Rfree (%)d | 20.2 | 21.6 | 21.1 |
| No. reflections in refinement | 56,957 | 79,920 | 64,194 |
| No. reflections in test set | 2932 | 4220 | 3443 |
| No. non-hydrogen atoms | |||
| Protein/substrate/solvent | 3518/—/457 | 7188/62/610 | 7262/46/16/650 |
| Mean B-factor (Å2) | |||
| Per monomer (A/B) | 13.20 | 22.15/22.21 | 17.69/17.20 |
| Per domain (A/B) | |||
| Domain 1 | 15.41 | 22.63/22.64 | 18.44/18.28 |
| Domain 2 | 10.00 | 18.68/19.54 | 15.28/15.10 |
| Domain 3 | 20.96 | 34.63/30.44 | 24.68/22.71 |
| ADS substrate (A/B) | — | 21.06/28.22 | — |
| AMP, FUM (A/B) | — | 12.66, 17.22/13.95, 16.91 | |
| C3 loop (A/B) | — | 24.30/41.16 | 20.30/18.56 |
| Solvent | 25.40 | 29.21 | 25.12 |
| rms deviation from ideal | |||
| Bond lengths (Å)/angles (°) | 0.005/1.21 | 0.006/1.31 | 0.005/1.23 |
| Dihedral/improper angles (°) | 19.61/0.71 | 19.77/0.80 | 19.68/0.73 |
| Ramachandran Plote | |||
| Total favored (%) | 92.3 | 91.3 | 91.3 |
| Total allowed (%) | 7.2 | 8.1 | 8.2 |
a
Rsym = Σ|I−<I>|/ΣI, where I is the measured intensity for symmetry-related reflections and <I> is the mean intensity for the reflection.
b
Last resolution shell extends from 2.07 to 2.00 and from 1.92–1.85 Å, respectively.
c
Rcrys = Σ(|Fo|−|Fc|)/Σ(|Fo|).
d
Rfree = Σ(|Fos|−|Fcs|)/Σ(|Fos|) where s refers to a subset of data not used in the refinement, representing 5% of the total number of observations.
e
According to the Ramachandran plot in PROCHECK.31