Rebinding of the extrinsic proteins of Photosystem II studied by electron microscopy and single particle alignment: an assessment with small two-dimensional ordered arrays of Photosystem II

Mark F. Rosenberg (Lead / Corresponding author), Andreas Holzenburg, Fiona H. Shepherd, William V Nicholson, Toby D. Flint, Robert C Ford

Research output: Contribution to journalArticle

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Abstract

Structural analysis of weakly ordered two-dimensional (2-D) ordered arrays of Photosystem II (a photosynthetic membrane protein complex) has been carried out using single-particle alignment and averaging (SPA). Although this averaging technique is normally applied to randomly oriented solubilised particles, we have found that it is also highly effective for the analysis of membrane-located 2-D ordered arrays and that the arrays can be used to check the effectiveness of the rotational alignment routines. Successful rotational alignment varied in the range 75 to 66% and 26 to 19%, depending on the stain used and hence the signal-to-noise ratio, and the data were used to address questions of space group assignments for Photosystem II 2-D ordered arrays. We thus hoped to settle the controversy as to the oligomeric form of PS II in native thylakoids, i.e., the monomeric vs. dimeric model. The methods were applied to the study of the removal and rebinding of extrinsic subunits in Photosystem II. Information on the location of the subunits in this large membrane protein complex is presented here.
Original languageEnglish
Pages (from-to)119-132
Number of pages14
JournalBiochimica et Biophysica Acta (BBA) - Bioenergetics
Volume1319
Issue number1
DOIs
Publication statusPublished - 28 Mar 1997

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Photosystem II Protein Complex
Electron microscopy
Electron Microscopy
Photosynthetic membranes
Membrane Proteins
Thylakoids
Proteins
Signal-To-Noise Ratio
Structural analysis
Signal to noise ratio
Coloring Agents
Membranes

Cite this

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title = "Rebinding of the extrinsic proteins of Photosystem II studied by electron microscopy and single particle alignment: an assessment with small two-dimensional ordered arrays of Photosystem II",
abstract = "Structural analysis of weakly ordered two-dimensional (2-D) ordered arrays of Photosystem II (a photosynthetic membrane protein complex) has been carried out using single-particle alignment and averaging (SPA). Although this averaging technique is normally applied to randomly oriented solubilised particles, we have found that it is also highly effective for the analysis of membrane-located 2-D ordered arrays and that the arrays can be used to check the effectiveness of the rotational alignment routines. Successful rotational alignment varied in the range 75 to 66{\%} and 26 to 19{\%}, depending on the stain used and hence the signal-to-noise ratio, and the data were used to address questions of space group assignments for Photosystem II 2-D ordered arrays. We thus hoped to settle the controversy as to the oligomeric form of PS II in native thylakoids, i.e., the monomeric vs. dimeric model. The methods were applied to the study of the removal and rebinding of extrinsic subunits in Photosystem II. Information on the location of the subunits in this large membrane protein complex is presented here.",
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Rebinding of the extrinsic proteins of Photosystem II studied by electron microscopy and single particle alignment: an assessment with small two-dimensional ordered arrays of Photosystem II. / Rosenberg, Mark F. (Lead / Corresponding author); Holzenburg, Andreas; Shepherd, Fiona H.; Nicholson, William V; Flint, Toby D.; Ford, Robert C.

In: Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1319, No. 1, 28.03.1997, p. 119-132.

Research output: Contribution to journalArticle

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AU - Nicholson, William V

AU - Flint, Toby D.

AU - Ford, Robert C

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AB - Structural analysis of weakly ordered two-dimensional (2-D) ordered arrays of Photosystem II (a photosynthetic membrane protein complex) has been carried out using single-particle alignment and averaging (SPA). Although this averaging technique is normally applied to randomly oriented solubilised particles, we have found that it is also highly effective for the analysis of membrane-located 2-D ordered arrays and that the arrays can be used to check the effectiveness of the rotational alignment routines. Successful rotational alignment varied in the range 75 to 66% and 26 to 19%, depending on the stain used and hence the signal-to-noise ratio, and the data were used to address questions of space group assignments for Photosystem II 2-D ordered arrays. We thus hoped to settle the controversy as to the oligomeric form of PS II in native thylakoids, i.e., the monomeric vs. dimeric model. The methods were applied to the study of the removal and rebinding of extrinsic subunits in Photosystem II. Information on the location of the subunits in this large membrane protein complex is presented here.

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