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. 2017 Mar 6;36(8):1084–1099. doi: 10.15252/embj.201696173

Figure EV4. Structure determination of the SCIN‐stabilized C3bBbPc complex.

Figure EV4

  1. Crystals of the complex (top), maximum dimensions are 100 × 20 × 5 μm3. An SDS–PAGE analysis of washed crystals with subsequent silver stain of the acrylamide gel (bottom) showed that all components are present.
  2. Crystal packing lattice for the complex. FPc is essential for lattice contacts, but presumably due to its pronounced non‐globular structure tight crystal packing is not possible resulting in solvent content of approx. 85% explaining the relative poor resolution of the diffraction data.
  3. Docking of six identical polyalanine TSRs in the real space averaged 2mFo‐DFc electron density. Two of the domains are tentatively assigned as TSR4 and TSR5 based on crystal packing constraints and functional data. The electron density map is calculated without any contribution from the docked TSRs.
  4. SEC on a 24 ml Superdex 200 Increase column with 0.72 nmol FPc (blue), 1.9 nmol C3 C345c domain (light blue) or combined (bark blue), performed in 10 mM HEPES pH 7.2, 75 mM NaCl.
  5. Relevant fractions from the combined run were analyzed by SDS–PAGE showing that FPc (25.5 + 23.9 kDa) and C3 C345c (17.4 kDa) did not co‐elute.