Open AccessPore dynamics and asymmetric cargo loading in an encapsulin nanocompartment
Author(s) -
Julia M. Ross,
Zak McIver,
Thomas Lambert,
Cecilia Piergentili,
Jasmine Emma Bird,
Kelly J. Gallagher,
Faye Cruickshank,
PW James,
Efrain Zarazúa-Arvizu,
Louise Horsfall,
Kevin J. Waldron,
Marcus D. Wilson,
C. Logan Mackay,
Arnaud Baslé,
David J. Clarke,
Jon MarlesWright
Publication year - 2022
Publication title -
science advances
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.928
H-Index - 146
ISSN - 2375-2548
DOI - 10.1126/sciadv.abj4461
Subject(s) - icosahedral symmetry , ferritin , chemistry , mass spectrometry , crystallography , biophysics , materials science , chemical engineering , chemical physics , nanotechnology , biochemistry , biology , chromatography , engineering
Encapsulins are protein nanocompartments that house various cargo enzymes, including a family of decameric ferritin-like proteins. Here, we study a recombinantHaliangium 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.