biostudies-literatureUnknown85(13)Rothemann RAApoptosis-inducing factor 1 (AIFM1) is a flavoprotein essential for mitochondrial function and biogenesis. Its interaction with MIA40/CHCHD4, the central component of the mitochondrial disulfide relay, accounts for some, but not all, aspects of AIFM1 function. We provide a high-confidence AIFM1 interactome that elucidates functional partners within the mitochondrial intermembrane space. We found that AIFM1 binding to adenylate kinase 2 (AK2), an essential enzyme that maintains cellular adenine nucleotide pools, depends on the AK2 C-terminal domain. High-resolution cryoelectron microscopy (cryo-EM) and biochemical analyses showed that both MIA40 and AK2A bind the AIFM1 C-terminal β-sheet domain. Their binding enhances NADH oxidoreductase activity by locking an active dimer conformation and,Molecular cell2550-2566.e6https://www.ebi.ac.uk/biostudies/studies/S-EPMC7617965biostudies-literatureInteraction with AK2A links AIFM1 to cellular energy metabolism.PMC7617965Riemer JMondal MStillger KPetrungaro CStobbe DNguyen THDMostert SRacho JRothemann RABano DGerlich SEhninger DLapacz KWeiss KNeundorf ISalscheider SLDengjel JGrobushkin PPoepsel SPavlenko EfalseInteraction with AK2A links AIFM1 to cellular energy metabolism.Apoptosis-inducing factor 1 (AIFM1) is a flavoprotein essential for mitochondrial function and biogenesis. Its interaction with MIA40/CHCHD4, the central component of the mitochondrial disulfide relay, accounts for some, but not all, aspects of AIFM1 function. We provide a high-confidence AIFM1 interactome that elucidates functional partners within the mitochondrial intermembrane space. We found that AIFM1 binding to adenylate kinase 2 (AK2), an essential enzyme that maintains cellular adenine nucleotide pools, depends on the AK2 C-terminal domain. High-resolution cryoelectron microscopy (cryo-EM) and biochemical analyses showed that both MIA40 and AK2A bind the AIFM1 C-terminal β-sheet domain. Their binding enhances NADH oxidoreductase activity by locking an active dimer conformation and,2025-01-01T00:00:00Z2025 Jul2026-03-31T10:36:54.812Z2025-08-24T03:06:55.162ZS-EPMC76179654057834810.1016/j.molcel.2025.05.036