8D4T

Mammalian CIV with GDN bound

Summary for 8D4T

• Entry DOI: 10.2210/pdb8d4t/pdb
• EMDB information: 27196
• Descriptor: Cytochrome c oxidase subunit 1, Cytochrome c oxidase subunit 7A1, mitochondrial, Cytochrome c oxidase subunit 7B, mitochondrial, ... (21 entities in total)
• Functional Keywords: mitochondrial electron transport chain, inhibitor, complex iv, membrane protein
• Biological source: Bos taurus (cattle); More
• Total number of polymer chains: 13
• Total formula weight: 202810.92

Authors

Di Trani, J.,Rubinstein, J. (deposition date: 2022-06-02, release date: 2022-07-06, Last modification date: 2024-12-25)

Primary citation

Di Trani, J.M.,Moe, A.,Riepl, D.,Saura, P.,Kaila, V.R.I.,Brzezinski, P.,Rubinstein, J.L.
Structural basis of mammalian complex IV inhibition by steroids.
Proc.Natl.Acad.Sci.USA, 119:e2205228119-e2205228119, 2022

PubMed Abstract: The mitochondrial electron transport chain maintains the proton motive force that powers adenosine triphosphate (ATP) synthesis. The energy for this process comes from oxidation of reduced nicotinamide adenine dinucleotide (NADH) and succinate, with the electrons from this oxidation passed via intermediate carriers to oxygen. Complex IV (CIV), the terminal oxidase, transfers electrons from the intermediate electron carrier cytochrome to oxygen, contributing to the proton motive force in the process. Within CIV, protons move through the K and D pathways during turnover. The former is responsible for transferring two protons to the enzyme's catalytic site upon its reduction, where they eventually combine with oxygen and electrons to form water. CIV is the main site for respiratory regulation, and although previous studies showed that steroid binding can regulate CIV activity, little is known about how this regulation occurs. Here, we characterize the interaction between CIV and steroids using a combination of kinetic experiments, structure determination, and molecular simulations. We show that molecules with a sterol moiety, such as glyco-diosgenin and cholesteryl hemisuccinate, reversibly inhibit CIV. Flash photolysis experiments probing the rapid equilibration of electrons within CIV demonstrate that binding of these molecules inhibits proton uptake through the K pathway. Single particle cryogenic electron microscopy (cryo-EM) of CIV with glyco-diosgenin reveals a previously undescribed steroid binding site adjacent to the K pathway, and molecular simulations suggest that the steroid binding modulates the conformational dynamics of key residues and proton transfer kinetics within this pathway. The binding pose of the sterol group sheds light on possible structural gating mechanisms in the CIV catalytic cycle.

PubMed: 35858451
DOI: 10.1073/pnas.2205228119

Experimental method

ELECTRON MICROSCOPY (3.1 Å)