Integrating on-grid immunogold labeling and cryo-electron tomography to reveal photosystem II structure and spatial distribution in thylakoid membranes.
Cryo-electron tomography (cryoET)
Immunogold labeling
Photosynthesis
Photosystem II (PSII)
Protein identification
Spatial distribution
Journal
Journal of structural biology
ISSN: 1095-8657
Titre abrégé: J Struct Biol
Pays: United States
ID NLM: 9011206
Informations de publication
Date de publication:
09 2021
09 2021
Historique:
received:
14
11
2020
revised:
10
05
2021
accepted:
13
05
2021
pubmed:
20
5
2021
medline:
5
4
2022
entrez:
19
5
2021
Statut:
ppublish
Résumé
A long-standing challenge in cell biology is elucidating the structure and spatial distribution of individual membrane-bound proteins, protein complexes and their interactions in their native environment. Here, we describe a workflow that combines on-grid immunogold labeling, followed by cryo-electron tomography (cryoET) imaging and structural analyses to identify and characterize the structure of photosystem II (PSII) complexes. Using an antibody specific to a core subunit of PSII, the D1 protein (uniquely found in the water splitting complex in all oxygenic photoautotrophs), we identified PSII complexes in biophysically active thylakoid membranes isolated from a model marine diatom Phaeodactylum tricornutum. Subsequent cryoET analyses of these protein complexes resolved two PSII structures: supercomplexes and dimeric cores. Our integrative approach establishes the structural signature of multimeric membrane protein complexes in their native environment and provides a pathway to elucidate their high-resolution structures.
Identifiants
pubmed: 34010667
pii: S1047-8477(21)00051-4
doi: 10.1016/j.jsb.2021.107746
pmc: PMC8577061
mid: NIHMS1707184
pii:
doi:
Substances chimiques
Light-Harvesting Protein Complexes
0
Photosystem II Protein Complex
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
107746Subventions
Organisme : NIGMS NIH HHS
ID : U24 GM116789
Pays : United States
Informations de copyright
Copyright © 2021 Elsevier Inc. All rights reserved.
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