Structural Basis of Membrane Protein Chaperoning through the Mitochondrial Intermembrane Space

The exchange of metabolites between the mitochondria matrix and the cytosol depends on beta-barrel channels in the outer membrane and alpha-helical carrier proteins in the inner membrane. The essential translocase of the inner membrane (TIM) chaperones escort these proteins through the intermembrane space, but the structural and mechanistic details remain elusive. We have used an integrated structural biology approach to reveal the functional principle of TIM chaperones. Multiple clamp-like binding sites hold the mitochondrial membrane proteins in a translocation-competent elongated form, thus mimicking characteristics of co-translational membrane insertion. The bound preprotein undergoes conformational dynamics within the chaperone binding clefts, pointing to a multitude of dynamic local binding events. Mutations in these binding sites cause cell death or growth defects associated with impairment of carrier and n-barrel protein biogenesis. Our work reveals how a single mitochondria! transfer-chaperone system is able to guide alpha-helical and beta-barrel membrane proteins in a nascent chain-like conformation through a ribosome-free compartment.