Cryo-EM structures of engineered active bc<sub>1</sub>-cbb<sub>3</sub> type CIII<sub>2</sub>CIV super-complexes and electronic communication between the complexes.

Steimle, Stefan; van Eeuwen, Trevor; Ozturk, Yavuz; Kim, Hee Jong; Braitbard, Merav; Selamoglu, Nur; Garcia, Benjamin A; Schneidman-Duhovny, Dina et al. · Nat Commun · 2021

basic_science · Level V

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Abstract

Respiratory electron transport complexes are organized as individual entities or combined as large supercomplexes (SC). Gram-negative bacteria deploy a mitochondrial-like cytochrome (cyt) bc<sub>1</sub> (Complex III, CIII<sub>2</sub>), and may have specific cbb<sub>3</sub>-type cyt c oxidases (Complex IV, CIV) instead of the canonical aa<sub>3</sub>-type CIV. Electron transfer between these complexes is mediated by soluble (c<sub>2</sub>) and membrane-anchored (c<sub>y</sub>) cyts. Here, we report the structure of an engineered bc<sub>1</sub>-cbb<sub>3</sub> type SC (CIII<sub>2</sub>CIV, 5.2 Å resolution) and three conformers of native CIII<sub>2</sub> (3.3 Å resolution). The SC is active in vivo and in vitro, contains all catalytic subunits and cofactors, and two extra transmembrane helices attributed to cyt c<sub>y</sub> and the assembly factor CcoH. The cyt c<sub>y</sub> is integral to SC, its cyt domain is mobile and it conveys electrons to CIV differently than cyt c<sub>2</sub>. The successful production of a native-like functional SC and determination of its structure illustrate the characteristics of membrane-confined and membrane-external respiratory electron transport pathways in Gram-negative bacteria.

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