Cryo-EM structures of engineered active bc 1-cbb 3 type CIII2CIV super-complexes and electronic communication between the complexes

Stefan Steimle; Trevor van Eeuwen; Yavuz Ozturk; Hee Jong Kim; Merav Braitbard; Nur Selamoglu; Benjamin A. Garcia; Dina Schneidman-Duhovny; Kenji Murakami; Fevzi Daldal

doi:https://doi.org/10.1038/s41467-021-21051-4

Cryo-EM structures of engineered active bc ₁-cbb ₃ type CIII₂CIV super-complexes and electronic communication between the complexes

Stefan Steimle, Trevor van Eeuwen, Yavuz Ozturk, Hee Jong Kim, Merav Braitbard, Nur Selamoglu, Benjamin A. Garcia, Dina Schneidman-Duhovny, Kenji Murakami, Fevzi Daldal

The Rachel and Selim Benin School of Engineering and Computer Science

The Hebrew University of Jerusalem

Research output: Contribution to journal › Article › peer-review

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) bc₁ (Complex III, CIII₂), and may have specific cbb₃-type cyt c oxidases (Complex IV, CIV) instead of the canonical aa₃-type CIV. Electron transfer between these complexes is mediated by soluble (c₂) and membrane-anchored (c_y) cyts. Here, we report the structure of an engineered bc₁-cbb₃ type SC (CIII₂CIV, 5.2 Å resolution) and three conformers of native CIII₂ (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_y and the assembly factor CcoH. The cyt c_y is integral to SC, its cyt domain is mobile and it conveys electrons to CIV differently than cyt c₂. 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.

Original language	English
Article number	929
Journal	Nature Communications
Volume	12
Issue number	1
DOIs	https://doi.org/10.1038/s41467-021-21051-4
State	Published - 10 Feb 2021

All Science Journal Classification (ASJC) codes

General Chemistry
General Biochemistry,Genetics and Molecular Biology
General Physics and Astronomy

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Steimle, S., van Eeuwen, T., Ozturk, Y., Kim, H. J., Braitbard, M., Selamoglu, N., Garcia, B. A., Schneidman-Duhovny, D., Murakami, K., & Daldal, F. (2021). Cryo-EM structures of engineered active bc ₁-cbb ₃ type CIII₂CIV super-complexes and electronic communication between the complexes. Nature Communications, 12(1), Article 929. https://doi.org/10.1038/s41467-021-21051-4

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title = "Cryo-EM structures of engineered active bc 1-cbb 3 type CIII2CIV super-complexes and electronic communication between the complexes",

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) bc1 (Complex III, CIII2), and may have specific cbb3-type cyt c oxidases (Complex IV, CIV) instead of the canonical aa3-type CIV. Electron transfer between these complexes is mediated by soluble (c2) and membrane-anchored (cy) cyts. Here, we report the structure of an engineered bc1-cbb3 type SC (CIII2CIV, 5.2 {\AA} resolution) and three conformers of native CIII2 (3.3 {\AA} resolution). The SC is active in vivo and in vitro, contains all catalytic subunits and cofactors, and two extra transmembrane helices attributed to cyt cy and the assembly factor CcoH. The cyt cy is integral to SC, its cyt domain is mobile and it conveys electrons to CIV differently than cyt c2. 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.",

author = "Stefan Steimle and {van Eeuwen}, Trevor and Yavuz Ozturk and Kim, {Hee Jong} and Merav Braitbard and Nur Selamoglu and Garcia, {Benjamin A.} and Dina Schneidman-Duhovny and Kenji Murakami and Fevzi Daldal",

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AU - Steimle, Stefan

AU - van Eeuwen, Trevor

AU - Ozturk, Yavuz

AU - Kim, Hee Jong

AU - Braitbard, Merav

AU - Selamoglu, Nur

AU - Garcia, Benjamin A.

AU - Schneidman-Duhovny, Dina

AU - Murakami, Kenji

AU - Daldal, Fevzi

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N2 - Respiratory electron transport complexes are organized as individual entities or combined as large supercomplexes (SC). Gram-negative bacteria deploy a mitochondrial-like cytochrome (cyt) bc1 (Complex III, CIII2), and may have specific cbb3-type cyt c oxidases (Complex IV, CIV) instead of the canonical aa3-type CIV. Electron transfer between these complexes is mediated by soluble (c2) and membrane-anchored (cy) cyts. Here, we report the structure of an engineered bc1-cbb3 type SC (CIII2CIV, 5.2 Å resolution) and three conformers of native CIII2 (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 cy and the assembly factor CcoH. The cyt cy is integral to SC, its cyt domain is mobile and it conveys electrons to CIV differently than cyt c2. 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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Cryo-EM structures of engineered active bc ₁-cbb ₃ type CIII₂CIV super-complexes and electronic communication between the complexes

Abstract

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