TY - JOUR T1 - Pore dynamics and asymmetric cargo loading in an encapsulin nanocompartment JF - bioRxiv DO - 10.1101/2021.04.15.439977 SP - 2021.04.15.439977 AU - Jennifer Ross AU - Zak McIver AU - Thomas Lambert AU - Cecilia Piergentili AU - Jasmine Emma Bird AU - Kelly J. Gallagher AU - Faye L. Cruickshank AU - Patrick James AU - Efrain ZarazĂșa-Arvizu AU - Louise E. Horsfall AU - Kevin J. Waldron AU - Marcus D. Wilson AU - C. Logan Mackay AU - Arnaud BaslĂ© AU - David J. Clarke AU - Jon Marles-Wright Y1 - 2021/01/01 UR - http://biorxiv.org/content/early/2021/10/06/2021.04.15.439977.abstract N2 - Encapsulins are protein nanocompartments that house various cargo enzymes, including a family of decameric ferritin-like proteins. Here, we study a recombinant Haliangium ochraceum encapsulin:encapsulated ferritin complex using electron cryo-microscopy and hydrogen/deuterium exchange mass spectrometry to gain insight into the structural relationship between the encapsulin shell and its protein cargo. An asymmetric single particle reconstruction reveals four encapsulated ferritin decamers in a tetrahedral arrangement within the encapsulin nanocompartment. This leads to a symmetry mismatch between the protein cargo and the icosahedral encapsulin shell. The encapsulated ferritin decamers are offset from the interior face of the encapsulin shell. Using HDX-MS, we observed dynamic behavior of the major five-fold pore in the encapsulin shell and show the pore opening via the movement of the encapsulin A-domain. These data will accelerate efforts to engineer the encapsulation of heterologous cargo proteins and to alter the permeability of the encapsulin shell via pore modifications.Teaser Cryo-EM and HDX-MS analysis of an encapsulin nanocompartment shows that the pores at the five-fold icosahedral vertex of the shell are flexible.Competing Interest StatementThe authors have declared no competing interest. ER -