Cryo-EM Structures of Respiratory bc1-cbb3 type CIII2CIV Super-complex and Electronic Communication Between the Complexes Stefan Steimle1, Trevor VanEeuwen2, Yavuz Ozturk1, #, Hee Jong Kim2, Merav Braitbard3, Nur Selamoglu1, Benjamin A. Garcia4, Dina Schneidman-Duhovny3, Kenji Murakami4,* and Fevzi Daldal1,* 1Department of Biology, University of Pennsylvania, Philadelphia, PA, 19104; 2Biochemistry and Molecular Biophysics Graduate Group, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104; #Institute of Biochemistry and Molecular Biology, Faculty of Medicine, Albert Ludwigs University of Freiburg, 79104 Freiburg, Germany; 3School of Computer Science and Engineering, Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, 9190401, Israel and 4Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104 Running title: Bacterial respiratory cytochrome bc1-cbb3 supercomplex Key words: cytochrome bc1 or Complex III; Cytochrome cbb3 oxidase or Complex IV; respiratory super-complex; electron carrier cytochrome c; membrane-anchored cytochrome cy; soluble cytochrome c2; Rhodobacter capsulatus; respiratory electron transport chain *Corresponding authors: Fevzi Daldal:
[email protected] Phone: +1 215 898-4394 Kenji Murakami:
[email protected] Phone: +1 215 573-1125 1 Methods Bacterial strains and growth conditions Bacterial strains and plasmids used are listed in Table S1. LB medium supplemented as appropriate with ampicillin, gentamicin, kanamycin or tetracycline at 100, 12, 50 or 12.5 μg/mL, respectively, was used for growing E. coli strains at 37 °C (Darrouzet and Daldal, 2002). R. capsulatus strains were grown chemoheterotrophically under semi-aerobic/dark conditions at 35 °C on enriched (MPYE) medium, supplemented as needed with gentamicin, kanamycin, spectinomycin, or tetracycline at 3, 10, 10 or 2.5 μg/mL, respectively (Davidson et al., 1992).