Table 2.
Data collection and refinement statistics.
| Mutant |
Y305A Yeast |
Y305F Yeast |
Y305F E. Coli |
|---|---|---|---|
| Expression system |
S. cerevisiae
|
S. cerevisiae
|
E. coli
|
| Space group |
P212121 |
P21 |
P21 |
| Cell constants (a, b, c: Å, β) |
138.91, 147.15, 233.76.90° |
96.29, 232.47, 104.31, 93.68° |
94.52, 232.62, 104.15, 91.51° |
| Number of observations |
476664 |
933972 |
692670 |
| Number of unique reflections |
134330 |
313044 |
217301 |
| Average multiplicity |
3.5 |
3.0 |
3.2 |
| Resolution limits (outer shell) |
30-2.5Å (2.6-2.5Å) |
100-1.90 (1.97-1.90) |
100-2.05Å (2.12-2.05Å) |
| Rmergea (%) |
7.2 (22.4) |
6.0 (46.3) |
6.7 (31.8) |
| Completeness (%) |
79.1 (41.0) |
87.4 (53.9) |
77.4 (51.9) |
| <I/σ(I)>b |
14.1 (3.1) |
17.2 (2.2) |
14.2 (3.3) |
| Resolution range for refinement (Å) |
30-2.5 |
6.0-1.9 |
40-2.05 |
| Reflections used for refinement |
134165 |
272421 |
205462 |
| Working set |
120767 |
258930 |
195210 |
| Test set |
13398 |
13491 |
10252 |
| Rcrystc (%) |
18.5 |
21.3 |
19.3 |
| Rfreed (%) |
19.8 |
21.7 |
20.0 |
| Number of non-hydrogen protein atomse |
5187 |
5198 |
5235 |
| Number of solvent molecules |
517 |
456 |
506 |
| Number of copper ionse |
1 |
1 |
1 |
| Number of phosphate Ions |
0 |
3 |
0 |
| Extent of protomer Resolved |
Pro18-Val672 |
Pro18-Val672 |
Ala15-Glu675 |
| RMSD bond lengths (Å) |
0.010 |
0.006 |
0.006 |
| RMSD bong angles (°) |
1.7 |
1.6 |
1.6 |
| Mean B factor (Å2) | |||
| Protein | 28.9 | 27.0 | 28.5 |
| Solvent | 35.4 | 31.8 | 37.0 |
| Copper ion |
31.2 |
23.1 |
26.2 |
| RMS ΔB (main chain-main chain) |
1.1 |
1.5 |
1.3 |
| RMS ΔB (main chain-side chain |
1.4 |
1.8 |
1.7 |
| RMS ΔB (side chain-side chain) |
2.1 |
3.2 |
2.9 |
| Ramachandran plot (%) | |||
| Allowed region | 99.4 | 99.6 | 99.3 |
| Generously allowed region | 0.4 | 0.2 | 0.5 |
| Disallowed region | 0.2 | 0.2 | 0.3 |
a
Rmerge=ΣhΣiI(h)-Ii(h)/ΣhΣiIi(h), where Ii(h) and I(h) are the ith and mean measurements of reflection h.
b
I/σ(I) is the average signal to noise ratio for merged reflection intensities.
c
R=ΣhFo-Fc/ΣhFo, where Fo and Fc are the observed and calculated structure factor amplitudes of reflection h.
d
Rfree is the test reflection data set, about 5-10 % selected randomly for cross validation during crystallographic refinement (Ref. 27).
e
Number of atoms in one independent protomer refined using strict NCS.