Eur. J. Inorg. Chem WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim, 2012 ISSN SUPPORTING INFORMATION

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1 Eur. J. Inorg. Chem WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim, 2012 ISSN SUPPORTING INFORMATION DOI: /ejic Title: Structural Insight into the Prolyl Hydroxylase PHD2: A Molecular Dynamics and DFT Study Author(s): Christian R. Wick, Harald Lanig, Christof M. Jäger, Nicolai Burzlaff,* Timothy Clark*

2 Figure S1.Time-averaged structures obtained after 500 ns of MD-simulation without restraints after the first 2 ns. Each structure represents a structural average over 100 ns, colour coded from blue (first 100 ns) to red (last 100 ns). Secondary structure elements known from the crystal structure are labelled for orientation. The average structures confirm the structural stability of the DSBH in solution and reveal the most flexible regions of the system. Possible novel secondary structure elements formed during the simulations are described in the text. The 17mer peptide fragment of CODD is labelled CODD when present and not distinguished from PHD2 by colour.

3 Table S1. The square of the summed up dot products (dot 2 ) of the first eigenvectors of each simulation was calculated in comparison to open.2og* (Values are scaled by factor 1000) Name dot 2 open.2og closed.2og* open.un open.un9* open.un9.codd open.un9*.codd open.2og.codd open.2og*.codd closed.2og.codd 0.756

4 Figure S2. RMSD values of the CODD peptide's C -atoms in closed.2og.codd plotted against time. Time-averaged structures are shown below for important regions.

5 Figure S3. Essential Dynamics of the MD Simulations containing UN9. The first Eigenvectors are projected on the reference structure (minimized startgeometry of open.2og) multiplied by their coefficients obtained from the projection of the eigenvectors on the corresponding trajectory.

6 Figure S4. RMSD values of the CODD peptide's C -atoms in open.un9.codd and open.un9*.codd plotted against time. Time-averaged structures are shown for important regions.

7 Figure S5. Optimized structures of the quintet states of the DFT model systems a, b and c. While the complete structure optimised with PW91 (purple) is shown, only the coordinating atoms are shown for B3PW91(green) and OLYP(grey). In model c, the structure optimised with OLYP is shown separately, because of its different tetrahedral coordination geometry. The initial coordinates are shown as grey shades.

8 Table S2: Distances observed for model b (Å). Atom numbering corresponds to that used in Figure 9. Method Fe-N 1 Fe-N 2 Fe-O 1 Fe-O 2 Fe-O 3 Fe-O 4[a] B3PW PW OLYP [a] O 4 corresponds to the second coordinating oxygen atom of the carboxylate trans to N 1.

9 Table S3. RMSD between experimental and theoretical values of the observed Fe-ligands bond lengths of the small DFT models a and c. The EXAFS data was taken from studies on DAOCS. [Fehler! Textmarke nicht definiert.] The X-Ray values include only the distances to the facial triad and the water molecule and were left out for model c, as no crystal structure of the aqua complex is available. Method RMSD EXAFS a RMSD X-Ray a RMSD EXAFS c B3PW PW OLYP

10 Table S4. Comparison of Iron-ligand bond length obtained with G(d,p) and TZP basis sets for model a quintet state. Method Fe-N 1 Fe-N 2 Fe-O 1 Fe-O 2 Fe-O 3 Fe-O 4 RMSD [b] NImag model a PW91/6-311+G(d,p) PW91/TZP [c] PW91/TZPfc [a] [c] OLYP/6-311+G(d,p) OLYP/TZP OLYP/TZPfc [a] [a] fc = frozen core option set to small. [b] RMSD calculated between every Fe-Ligand bond for each density functional (calculation with the G(d,p) basis set used as reference.) [c] due to technical reasons only numerical frequencies were available for PW91 and TZP basis sets. Visual inspection of the imaginary frequencies clearly shows that they can be attributed to minima vibrations (Figure S7). All other frequencies could be obtained analytically.

11 Figure S6: Crystallographic B-factor plotted on the crystal structures 2HBT (top) and 3HQR (bottom) color-coded from green (< 30 Å 2 ) to white (30 Å 2 ) to red (> 30 Å 2 ).

12 Figure S7: Visualization of numerically obtained imaginary frequencies for geometry optimizations of model a with PW91/TZP (a), cm -1 ) and PW91/TZPfc (b), cm -1 ).