a. The PAS-GAF (blue and green, respectively) and the PSM (white)
constructs from D. radiodurans BphP in their dark-adapted Pr
states are shown superimposed. b. Superposition of atomic models of
the PSM in the room-temperature Pr state (colored) with that derived from
diffraction data collected at 100 K (white, PDB ID 4Q0J). The β-sheets
of the GAF domains were superposed, allowing the respective positions of the PAS
and PHY domains of the two models to be identified. The largest differences
between the models were found at the PHY domain. For the room temperature model
the domains were colored blue, green, and orange for the PAS, GAF, and PHY
domains, respectively. Several important features are labeled: HP, hairpin; NTE,
amino-terminal extension; BV, biliverdin. BV from the room temperature structure
is shown for orientation. c and d. Composite
simulated-annealing omit maps (2Fo-Fc) contoured at 1
σ were superimposed with the corresponding PAS-GAF (c) or
PSM (d) model of DrBphP22. For clarity only electron density around Cys24 (gray),
biliverdin (cyan) and for (c) the pyrrole water is shown.
e. Metal site electron density (2Fo-Fc)
at the heterodinuclear Mn/Fe site in aerobic class Ic RNR, metal ions and
protein ligands are indicated (contoured to 1.3 σ) in blue. Residual
positive difference electron density map (Fo-Fc)
representing non-protein ligands is indicated in green (contoured to 3.5
σ). The manganese and iron atoms are depicted as purple and orange
spheres, respectively. Kβ1,3 XES of oxidized RNR in crystal
and in solution collected at room temperature with an XFEL is shown to the left.
A Mn(II)Cl2 calibration standard is also shown to illustrate the
absolute oxidation state of the solution and crystal spectra. Fe Kα XES
Data collected from oxidized RNR crystals is shown at the right.