TABLE 1.
Data collection and refinement statistics
Dataset nomenclature is as follows, SeMet(L48M)Mub-R5: selenomethionyl derivative of the L48 M mutant of Mub-R5. Numbers in parentheses refer to data in the highest resolution bin.
| Dataset | Se-Met(L48M)Mub-R5 (SAD data collection) | Native Mub-R5 |
|---|---|---|
| Beamline | Diamond ID02 | ESRF BM14 |
| Space group | P212121 | P212121 |
| Cell parameters, a, b, c (Å) | 44.9, 45.7, 191.4 | 44.9, 45.8, 191.6 |
| Wavelength (Å) | 0.979 | 1.033 |
| Resolution (Å) | 60-2.0 (2.11-2.00) | 60-1.80 (1.90-1.80) |
| Rsyma (%) | 5.9 (9.6) | 6.0 (20.4) |
| Ranomb (%) | 5.2 (6.4) | |
| 〈I/σI〉 | 34.6 (25.0) | 13.8 (9.5) |
| Independent reflections | 27,636 (3,931) | 36,236 (4391) |
| Completeness (%) | 100.0 (100.0) | 96.0 (81.9) |
| Multiplicity | 14.0 (14.4) | 2.8 (1.9) |
| Anomalous completeness (%) | 100.0 (100.0) | |
| Anomalous multiplicity | 7.5 (7.5) | |
| Overall temperature factor (Å2) | 14.3 | 15.4 |
| FOMc (SAD phasing) | 0.82 (0.75) | |
| FOMc (solvent flattened) | 0.90 (0.82) | |
| Refinement statistics | ||
| Mub-R5 heterodimers per AUd | 2 | |
| Refined structure | ||
| Total atoms | 3,639 | |
| Water molecules | 755 | |
| Rcryste | 20.2 (26.9) | |
| Rfreef | 25.9 (30.6) | |
| Ramachandran analysisg (%) | ||
| Most favored | 98.9 | |
| Disallowed | 0.0 | |
| r.m.s.d. | ||
| Bonds (Å) | 0.009 | |
| Angles (°) | 1.16 | |
| Planes (Å) | 0.005 | |
| Mean atomic B-value (Å2) | 14.6 | |
a Rsym = Σ|Ii − 〈I〉|/Σ Ii, where 〈I〉 is the average of symmetry equivalent reflections, and the summation extends over all observations for all unique reflections.
b Ranom = Σ|〈I+〉 − 〈I−〉|/Σ|〈I+〉 + 〈I−〉| (reported for the selenium SAD dataset to a maximum resolution of 2.0 Å).
c Average figures of merit (FOM) after solvent flattening with RESOLVE (Terwilliger (36)).
d AU refers to the crystallographic asymmetric unit.
e Rcryst = Σ½|Fo| − |Fc½|/Σ|Fo|, where Fo and Fc are the measured and calculated structure factors, respectively.
f For Rfree the summations extends over a randomly selected subset (5%) of reflections excluded from all stages of refinement.
g Structure validation was performed using MOLPROBITY (83).