Safari with an Electron Gun: Visualization of Protein and Membrane Interactions in Mitochondria in Natural Environment

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Semen V. Nesterov^1,a*, Konstantin S. Plokhikh¹, Yuriy M. Chesnokov¹, Denis A. Mustafin¹, Tatyana N. Goleva¹, Anton G. Rogov¹, Raif G. Vasilov¹, and Lev S. Yaguzhinsky²

¹National Research Center “Kurchatov Institute,” 123182 Moscow, Russia

²Belozersky Research Institute for Physico-Chemical Biology, Lomonosov Moscow State University, 119992 Moscow, Russia

Received September 24, 2023; Revised February 6, 2024; Accepted February 7, 2024

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This paper presents new structural data about mitochondria using correlative light and electron microscopy (CLEM) and cryo-electron tomography. These state-of-the-art structural biology methods allow studying biological objects at nanometer scales under natural conditions. Non-invasiveness of these methods makes them comparable to observing animals in their natural environment on a safari. The paper highlights two areas of research that can only be accomplished using these methods. The study visualized location of the Aβ42 amyloid aggregates in relation to mitochondria to test a hypothesis of development of mitochondrial dysfunction in Alzheimer’s disease. The results showed that the Aβ42 aggregates do not interact with mitochondria, although some of them are closely located. Therefore, the study demonstrated that mitochondrial dysfunction is not directly associated with the effects of aggregates on mitochondrial structure. Other processes should be considered as sources of mitochondrial dysfunction. Second unique area presented in this work is high-resolution visualization of the mitochondrial membranes and proteins in them. Analysis of the cryo-ET data reveals toroidal holes in the lamellar structures of cardiac mitochondrial cristae, where ATP synthases are located. The study proposes a new mechanism for sorting and clustering protein complexes in the membrane based on topology. According to this suggestion, position of the OXPHOS system proteins in the membrane is determined by its curvature. High-resolution tomography expands and complements existing ideas about the structural and functional organization of mitochondria. This makes it possible to study the previously inaccessible structural interactions of proteins with each other and with membranes in vivo.

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KEY WORDS: membrane, mitochondria, oxidative phosphorylation, cryo-electron microscopy, supercomplex, ATP synthase, respirasome, Aβ42, amyloid aggregates

DOI: 10.1134/S0006297924020068

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