RT Journal Article SR Electronic T1 Transient protein structure guides surface diffusion pathways for electron transport in membrane supercomplexes JF bioRxiv FD Cold Spring Harbor Laboratory SP 2024.09.30.615558 DO 10.1101/2024.09.30.615558 A1 Chan, Chun Kit A1 Nguyen, Jonathan A1 Hryc, Corey F. A1 Gupta, Chitrak A1 Redding, Kevin A1 Dowhan, William A1 Baker, Matthew L. A1 Perez, Alberto A1 Mileykovskaya, Eugenia A1 Singharoy, Abhishek YR 2024 UL http://biorxiv.org/content/early/2024/10/01/2024.09.30.615558.abstract AB The biological significance of protein supercomplexes have remained contentious, particularly how they tune the shuttling of charge-carrier redox proteins across cell membranes during biological energy conversion. We employ multiscale modeling and single particle cryo-electron microscopy (cryo-EM) to determine the mechanisms of diffusive electron transfer in mitochondrial supercomplexes, composed of respiratory complexes III and IV (CIII and CIV). Using a combination of bioinformatic and entropy maximization tools, we model an ensemble of structures representing the conformational space of CIII’s disordered QCR6 ‘hinge’ within the yeast CIII2CIV2 supercomplex. Molecular and Brownian Dynamics simulations of the entire supercomplex reveal a mechanism for electrostatic coupling between these negatively charged hinge conformations, and binding and directional diffusion of the redox proteins on the mitochondrial membrane, which is simulated over the millisecond timescale. Anionic lipids reinforce this conformationally-coupled recognition of the supercomplex by retaining a pool of the redox proteins in the vicinity of the membrane when the hinges are of a critical length. Cryo-EM models reveal a large-scale rearrangement of the ΔQCR6 supercomplex, which retains a surprisingly robust electrostatic environment for recognition of the redox protein, despite compromise in the supercomplex’s negative charge, still enabling a surface-mediated electron transfer in this CIII2CIV2 variant. Altogether, the evolutionary need of confining electron carriers on the surface of bioenergetic membranes is found to give rise to a refolding-guided diffusion model of the redox proteins, which improves the energy conversion efficiency within the supercomplex by nearly 30%.Competing Interest StatementThe authors have declared no competing interest.