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TitleIntegrating on-grid immunogold labeling and cryo-electron tomography to reveal photosystem II structure and spatial distribution in thylakoid membranes.
Journal, issue, pagesJ Struct Biol, Vol. 213, Issue 3, Page 107746, Year 2021
Publish dateMay 16, 2021
AuthorsJennifer Jiang / Kuan Yu Cheong / Paul G Falkowski / Wei Dai /
PubMed AbstractA 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 ...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.
External linksJ Struct Biol / PubMed:34010667 / PubMed Central
MethodsEM (subtomogram averaging)
Resolution37.0 Å
Structure data

EMDB-23791:
Subtomogram average of Photosystem II 2D semicrystalline array on thylakoid membranes isolated from Phaeodactylum tricornutum
Method: EM (subtomogram averaging) / Resolution: 37.0 Å

Source
  • Phaeodactylum tricornutum CCAP 1055/1 (Diatom)

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