Functional role of respiratory supercomplexes in mice: SCAF1 relevance and segmentation of the Qpool

Mitochondrial respiratory complexes assemble into supercomplexes (SC). Q-respirasome (III2 + IV) requires the supercomplex assembly factor (SCAF1) protein. The role of this factor in the N-respirasome (I + III2 + IV) and the physiological role of SCs are controversial. Here, we study C57BL/6J mice harboring nonfunctional SCAF1, the full knockout for SCAF1, or the wild-type version of the protein and found that exercise performance is SCAF1 dependent. By combining quantitative data–independent proteomics, 2D Blue native gel electrophoresis, and functional analysis of enriched respirasome fractions, we show that SCAF1 confers structural attachment between III2 and IV within the N-respirasome, increases NADH-dependent respiration, and reduces reactive oxygen species (ROS). Furthermore, the expression of AOX in cells and mice confirms that CI-CIII superassembly segments the CoQ in two pools and modulates CI-NADH oxidative capacity.This study was supported by MINECO (SAF2015-65633-R), MCIU (RTI2018-099357-B-I00), CIBERFES (CB16/10/00282), and HFSP (RGP0016/2018) to J.A.E.; MINECO-BIO2015-67580-P and PGC2018-097019-B-I00, ISCIIIIPT13/0001 (ISCIII-SGEFI/FEDER, ProteoRed) the Fundació MaratóTV3 (grant 122/C/2015), and “la Caixa” Banking Foundation (project code HR17-00247) to J.V. The CNIC is supported by the Ministry of Economy, Industry and Competitiveness (MEIC) and the Pro-CNIC Foundation and is a Severo Ochoa Center of Excellence (MINECO award SEV-2015-0505).S