Proteomics

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Proteomic comparison of photosynthetic reaction centre-light harvesting 1 complexes in wild-type, protein-W knockout and His-tagged protein-W strains of Rhodopseudomonas palustris.


ABSTRACT: Rhodopseudomonas palustris is a species of purple phototrophic bacterium. In these species, solar energy is captured by reaction centre-light harvesting 1 (RC-LH1) complexes which reside in membranes within the cells. The RC comprises three protein subunits: H, M, L and is encircled by a ring of LH1-α and LH1-β subunit pairs. Approximately 10% of these complexes in the wild-type (WT) strain include an additional subunit called protein-W. In this project, we have determined cryo-EM structures for RC-LH1 complexes both with and without protein-W. To enable the purification of RC-LH1-W with 100% occupancy of protein-W, strain expressing a C-terminally His-tagged W was engineered. For complexes lacking W, a knockout strain was used. Proteomic analysis was employed to validate these strains and establish that the WT and W-His expressed similar levels of W relative to RC-LH1.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Rhodopseudomonas Palustris Cga009

SUBMITTER: Philip Jackson  

LAB HEAD: Christopher Neil Hunter

PROVIDER: PXD020402 | Pride | 2021-09-09

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
20191118_PJ_DS_W-His-3-1.raw Raw
20191118_PJ_DS_W-His-3-2.raw Raw
20191118_PJ_DS_W-His-3-3.raw Raw
20191118_PJ_DS_delta-W-2-1.raw Raw
20191118_PJ_DS_delta-W-2-2.raw Raw
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Publications


The reaction-center light-harvesting complex 1 (RC-LH1) is the core photosynthetic component in purple phototrophic bacteria. We present two cryo-electron microscopy structures of RC-LH1 complexes from <i>Rhodopseudomonas palustris</i> A 2.65-Å resolution structure of the RC-LH1<sub>14</sub>-W complex consists of an open 14-subunit LH1 ring surrounding the RC interrupted by protein-W, whereas the complex without protein-W at 2.80-Å resolution comprises an RC completely encircled by a closed, 16-  ...[more]

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