Cryo-EM of native membranes reveals an intimate connection between the Krebs cycle and respiration in mycobacteria

Avatar
Poster
Voice is AI-generated
Connected to paperThis paper is a preprint and has not been certified by peer review

Cryo-EM of native membranes reveals an intimate connection between the Krebs cycle and respiration in mycobacteria

Authors

Di Trani, J. M.; Yu, J.; Courbon, G. M.; Rodriguez, A. P. L.; Cheung, C.-Y.; Liang, Y.; Coupland, C. E.; Bueler, S. A.; Cook, G. M.; Brzezinski, P.; Rubinstein, J. L.

Abstract

Imaging of endogenous protein complexes in their native membranes can reveal protein-protein interactions that are lost upon detergent solubilization. To investigate interactions in the mycobacterial oxidative phosphorylation machinery, we prepared inverted membrane vesicles from Mycobacterium smegmatis and enriched for vesicles containing complexes of interest by affinity chromatography. Electron cryomicroscopy (cryo-EM) of these vesicles revealed that malate-quinone oxidoreductase (Mqo), an enzyme from the Krebs cycle, physically associates with the electron transport chain Complex III2IV2 (CIII2CIV2) supercomplex. Analysis of the Mqo:CIII2CIV2 interaction shows that CIII2CIV2 is necessary for malate-driven, but not NADH-driven, electron transport chain activity and oxygen consumption. Further, the association of Mqo with CIII2CIV2 enables electron transfer from malate to CIII2CIV2 with millisecond kinetics. Together, these findings indicate a connection between the Krebs cycle and respiration that directs electrons along a single branch of the mycobacterial electron transport chain.

Follow Us on

0 comments

Add comment