β-barrel proteins are present in the outer membrane of Gram-negative bacteria and of organel- les of endosymbiotic origin, i.e. mitochondria and chloroplasts where they perform a variety of functions. Mitochondrial β-barrel proteins are important for protein import, metabolite transport and the organelle morphology and distribution. They also seem to play a crucial role in mitochondria evolution. Quite recently a specific pathway for the insertion of β-barrel proteins was identified in both mitochondria and Gram-negative bacteria and was proved to be conserved during evolution.