Higher-order structural organization of mitochondrial metabolism

The mitochondrion embeds numerous metabolic, biosynthetic, and signaling pathways occurring within its specialized compartments that collectively define eukaryotic biology. To visualize these, cryogenic electron microscopy (cryo-EM) is now utilized, particularly for resolving biomolecular complexes that were notoriously hard to study with other structural methods. Our review synthesizes recent cryo-EM-based discoveries, analyzes structures related to mitochondrial metabolism, and explains related functional aspects of core energy metabolism, biosynthetic processes, transport, and regulatory systems. We report on recent data on mitochondrial enzyme filamentation, higher-order metabolons, enzyme-gating mechanisms, and contact-site nanostructures. In addition to single-particle cryo-EM results, we discuss emerging in situ cryo-tomography data, the integration of traditional structural biology approaches, and the use of in silico models for less-studied pathways. By mapping hundreds of recent mitochondrial structures, we provide a roadmap that connects structural biochemistry with cell physiology, disclosing the molecular basis of mitochondrial disease. Our collected resources may guide future integrative research aiming to elucidate mitochondrial architecture and function across organisms and conditions.