Comparison of photosystem II 3D structure as determined by electron crystallography of frozen-hydrated and negatively stained specimens

Svetla S. Stoylova, Toby D. Flint, Ashraf Kitmitto, Robert C. Ford, Andreas Holzenburg

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The efficiency of electron crystallographic analysis for 3D structure determination has been demonstrated for an increasing number of proteins and protein complexes. Here, we report cryo-electron crystallography data leading to the first 3D structure of unstained, frozen-hydrated photosystem II (PSII), a multi-protein, membrane-integral complex comprised of more than 20 polypeptides. These data are directly comparable with the 3D structure of negatively stained PSII and reveal a high correlation between the two types of specimens at 3 nm resolution, confirming the earlier assignments of subunits to well-defined domains as well as the functionally important intramolecular cavity. The cavity is at least partially capped on the lumenal surface of the complex and opens out to a 3-nm diameter internal water-filled space that dominates the lumenal side of PSII. The data are discussed in the light of recent studies on PSII subcomplexes. It is concluded that the fidelity of data on transmembrane proteins in negative stain is mainly affected by a flattening of the specimen, partial-depth staining and limited stain penetration into the lipid bilayer.

Original languageEnglish (US)
Pages (from-to)341-348
Number of pages8
JournalMicron
Volume29
Issue number5
DOIs
StatePublished - Oct 1998
Externally publishedYes

Fingerprint

Crystallography
Photosystem II Protein Complex
crystallography
Electrons
proteins
Proteins
electrons
Coloring Agents
cavities
Lipid bilayers
Polypeptides
polypeptides
Lipid Bilayers
flattening
staining
lipids
Membrane Proteins
penetration
Staining and Labeling
membranes

Keywords

  • 2D crystals
  • 3D structure
  • Barley
  • Cryo-electron crystallography
  • Frozen-hydrated
  • Negative staining
  • Photosystem II

ASJC Scopus subject areas

  • Cell Biology
  • Materials Science(all)
  • Instrumentation

Cite this

Comparison of photosystem II 3D structure as determined by electron crystallography of frozen-hydrated and negatively stained specimens. / Stoylova, Svetla S.; Flint, Toby D.; Kitmitto, Ashraf; Ford, Robert C.; Holzenburg, Andreas.

In: Micron, Vol. 29, No. 5, 10.1998, p. 341-348.

Research output: Contribution to journalArticle

Stoylova, Svetla S. ; Flint, Toby D. ; Kitmitto, Ashraf ; Ford, Robert C. ; Holzenburg, Andreas. / Comparison of photosystem II 3D structure as determined by electron crystallography of frozen-hydrated and negatively stained specimens. In: Micron. 1998 ; Vol. 29, No. 5. pp. 341-348.
@article{02d7520b24dd41e59ea200b3bf4e4025,
title = "Comparison of photosystem II 3D structure as determined by electron crystallography of frozen-hydrated and negatively stained specimens",
abstract = "The efficiency of electron crystallographic analysis for 3D structure determination has been demonstrated for an increasing number of proteins and protein complexes. Here, we report cryo-electron crystallography data leading to the first 3D structure of unstained, frozen-hydrated photosystem II (PSII), a multi-protein, membrane-integral complex comprised of more than 20 polypeptides. These data are directly comparable with the 3D structure of negatively stained PSII and reveal a high correlation between the two types of specimens at 3 nm resolution, confirming the earlier assignments of subunits to well-defined domains as well as the functionally important intramolecular cavity. The cavity is at least partially capped on the lumenal surface of the complex and opens out to a 3-nm diameter internal water-filled space that dominates the lumenal side of PSII. The data are discussed in the light of recent studies on PSII subcomplexes. It is concluded that the fidelity of data on transmembrane proteins in negative stain is mainly affected by a flattening of the specimen, partial-depth staining and limited stain penetration into the lipid bilayer.",
keywords = "2D crystals, 3D structure, Barley, Cryo-electron crystallography, Frozen-hydrated, Negative staining, Photosystem II",
author = "Stoylova, {Svetla S.} and Flint, {Toby D.} and Ashraf Kitmitto and Ford, {Robert C.} and Andreas Holzenburg",
year = "1998",
month = "10",
doi = "10.1016/S0968-4328(98)00017-1",
language = "English (US)",
volume = "29",
pages = "341--348",
journal = "Micron",
issn = "0968-4328",
publisher = "Elsevier Limited",
number = "5",

}

TY - JOUR

T1 - Comparison of photosystem II 3D structure as determined by electron crystallography of frozen-hydrated and negatively stained specimens

AU - Stoylova, Svetla S.

AU - Flint, Toby D.

AU - Kitmitto, Ashraf

AU - Ford, Robert C.

AU - Holzenburg, Andreas

PY - 1998/10

Y1 - 1998/10

N2 - The efficiency of electron crystallographic analysis for 3D structure determination has been demonstrated for an increasing number of proteins and protein complexes. Here, we report cryo-electron crystallography data leading to the first 3D structure of unstained, frozen-hydrated photosystem II (PSII), a multi-protein, membrane-integral complex comprised of more than 20 polypeptides. These data are directly comparable with the 3D structure of negatively stained PSII and reveal a high correlation between the two types of specimens at 3 nm resolution, confirming the earlier assignments of subunits to well-defined domains as well as the functionally important intramolecular cavity. The cavity is at least partially capped on the lumenal surface of the complex and opens out to a 3-nm diameter internal water-filled space that dominates the lumenal side of PSII. The data are discussed in the light of recent studies on PSII subcomplexes. It is concluded that the fidelity of data on transmembrane proteins in negative stain is mainly affected by a flattening of the specimen, partial-depth staining and limited stain penetration into the lipid bilayer.

AB - The efficiency of electron crystallographic analysis for 3D structure determination has been demonstrated for an increasing number of proteins and protein complexes. Here, we report cryo-electron crystallography data leading to the first 3D structure of unstained, frozen-hydrated photosystem II (PSII), a multi-protein, membrane-integral complex comprised of more than 20 polypeptides. These data are directly comparable with the 3D structure of negatively stained PSII and reveal a high correlation between the two types of specimens at 3 nm resolution, confirming the earlier assignments of subunits to well-defined domains as well as the functionally important intramolecular cavity. The cavity is at least partially capped on the lumenal surface of the complex and opens out to a 3-nm diameter internal water-filled space that dominates the lumenal side of PSII. The data are discussed in the light of recent studies on PSII subcomplexes. It is concluded that the fidelity of data on transmembrane proteins in negative stain is mainly affected by a flattening of the specimen, partial-depth staining and limited stain penetration into the lipid bilayer.

KW - 2D crystals

KW - 3D structure

KW - Barley

KW - Cryo-electron crystallography

KW - Frozen-hydrated

KW - Negative staining

KW - Photosystem II

UR - http://www.scopus.com/inward/record.url?scp=0032192445&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032192445&partnerID=8YFLogxK

U2 - 10.1016/S0968-4328(98)00017-1

DO - 10.1016/S0968-4328(98)00017-1

M3 - Article

AN - SCOPUS:0032192445

VL - 29

SP - 341

EP - 348

JO - Micron

JF - Micron

SN - 0968-4328

IS - 5

ER -