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Pore dynamics and asymmetric cargo loading in an encapsulin nanocompartment.

Lookup NU author(s): Dr Jenn RossORCiD, Dr Zak McIverORCiD, Dr Cecilia Piergentili, Dr Jasmine Bird, Dr Kevin WaldronORCiD, Dr Arnaud Basle, Dr Jon Marles-WrightORCiD

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).

Abstract

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 cryo–electron 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 hydrogen/deuterium exchange mass spectrometry, we observed the dynamic behavior of the major fivefold 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.

Publication metadata

Author(s): Ross J, McIver Z, Lambert T, Piergentili C, Bird JE, Gallagher KJ, Cruickshank FL, James P, Zarazúa-Arvizu E, Horsfall LE, Waldron KJ, Wilson MD, Mackay CL, Baslé A, Clarke DJ, Marles-Wright J

Publication type: Article

Publication status: Published

Journal: Science Advances

Year: 2022

Volume: 8

Issue: 4

Online publication date: 26/01/2022

Acceptance date: 02/12/2021

Date deposited: 08/02/2022

ISSN (electronic): 2375-2548

Publisher: AAAS

URL: https://doi.org/10.1126/sciadv.abj4461

DOI: 10.1126/sciadv.abj4461

PubMed id: 35080974

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