Proteomics

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In-cell structures of a conserved supramolecular array at the mitochondria-cytoskeleton interface in mammalian sperm


ABSTRACT: Mitochondria-cytoskeleton interactions modulate cellular physiology by regulating mitochondrial transport, positioning, and immobilization. However, there is very little structural information defining mitochondria-cytoskeleton interfaces in any cell type. Here, we use cryo-focused ion beam milling-enabled cryo-electron tomography to image mammalian sperm, where mitochondria wrap around the ciliary cytoskeleton. We find that mitochondria are tethered to their neighbors through inter-mitochondrial linkers and are anchored to the cytoskeleton through ordered arrays on the outer mitochondrial membrane. We use subtomogram averaging to resolve in-cell structures of these arrays from three mammalian species, revealing they are conserved across species despite variations in mitochondrial dimensions and cristae organization. We find that the arrays consist of boat-shaped particles anchored on a network of membrane pores whose arrangement and dimensions are consistent with voltage dependent anion channels. Proteomics and in-cell cross-linking mass spectrometry suggest that the conserved arrays are composed of glycerol kinase-like proteins. Ordered supramolecular assemblies may serve to stabilize similar contact sites in other cell types where mitochondria need to be immobilized in specific subcellular environments, such as in muscles and neurons.

INSTRUMENT(S): Orbitrap Fusion Lumos, Q Exactive HF

ORGANISM(S): Sus Scrofa Domesticus (domestic Pig)

TISSUE(S): Spermatozoon

SUBMITTER: Riccardo Zenezini Chiozzi  

LAB HEAD: Albert Heck

PROVIDER: PXD025562 | Pride | 2021-10-29

REPOSITORIES: Pride

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Publications

In-cell structures of conserved supramolecular protein arrays at the mitochondria-cytoskeleton interface in mammalian sperm.

Leung Miguel Ricardo MR   Zenezini Chiozzi Riccardo R   Roelofs Marc C MC   Hevler Johannes F JF   Ravi Ravi Teja RT   Maitan Paula P   Zhang Min M   Henning Heiko H   Bromfield Elizabeth G EG   Howes Stuart C SC   Gadella Bart M BM   Heck Albert J R AJR   Zeev-Ben-Mordehai Tzviya T  

Proceedings of the National Academy of Sciences of the United States of America 20211101 45


Mitochondria-cytoskeleton interactions modulate cellular physiology by regulating mitochondrial transport, positioning, and immobilization. However, there is very little structural information defining mitochondria-cytoskeleton interfaces in any cell type. Here, we use cryofocused ion beam milling-enabled cryoelectron tomography to image mammalian sperm, where mitochondria wrap around the flagellar cytoskeleton. We find that mitochondria are tethered to their neighbors through intermitochondrial  ...[more]

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