DOI: 10.1111/j.0022-2720.2004.01395.x. Direct visualization of receptor arrays in frozen‐hydrated sections and plunge‐frozen specimens of <i>E. coli</i> engineered to overproduce the chemotaxis receptor Tsr

Direct visualization of receptor arrays in frozen‐hydrated sections and plunge‐frozen specimens of E. coli engineered to overproduce the chemotaxis receptor Tsr

10.1111/j.0022-2720.2004.01395.x

Crossref journal-article

Wiley

Journal of Microscopy (311)

Abstract

SummaryWe have recently reported electron tomographic studies of sections obtained from chemically fixed E. coli cells overproducing the 60‐kDa chemotaxis receptor Tsr. Membrane extracts from these cells prepared in the presence of Tween‐80 display hexagonally close‐packed microcrystalline assemblies of Tsr, with a repeating unit large enough to accommodate six Tsr molecules arranged as trimers of receptor dimers. Here, we report the direct visualization of the Tsr receptor clusters in (i) vitrified cell suspensions of cells overproducing Tsr, prepared by rapid plunge‐freezing, and (ii) frozen‐hydrated sections obtained from cells frozen under high pressure. The frozen‐hydrated sections were generated by sectioning at −150 °C using a diamond knife with a 25° knife angle, with nominal thicknesses ranging from 20 to 60 nm. There is excellent correspondence between the spatial arrangement of receptors in thin frozen‐hydrated sections and the arrangements found in negatively stained membrane extracts and plunge‐frozen cells, highlighting the potential of using frozen‐hydrated sections for the study of macromolecular assemblies within cells under near‐native conditions.

Bibliography

ZHANG, P., BOS, E., HEYMANN, J., GNAEGI, H., KESSEL, M., PETERS, P. J., & SUBRAMANIAM, S. (2004). Direct visualization of receptor arrays in frozenâhydrated sections and plungeâfrozen specimens of E.Â coli engineered to overproduce the chemotaxis receptor Tsr. Journal of Microscopy, 216(1), 76â83. Portico.

Authors 7

P. ZHANG (first)
E. BOS (additional)
J. HEYMANN (additional)
H. GNAEGI (additional)
M. KESSEL (additional)
P. J. PETERS (additional)
S. SUBRAMANIAM (additional)

References 30 Referenced 65

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{'issue': '2', 'key': 'e_1_2_5_30_1', 'first-page': '1238', 'article-title': 'Ultramicrotomy of polymers using an oscillating knife; improving polymer morphology', 'volume': '8', 'author': 'Vastenhout J.S.', 'year': '2002', 'journal-title': 'Microsc. Microanal'} / Microsc. Microanal / Ultramicrotomy of polymers using an oscillating knife; improving polymer morphology by Vastenhout J.S. (2002)
10.1128/JB.185.12.3636-3643.2003

Dates

Type	When
Created	20 years, 11 months ago (Sept. 15, 2004, 11:46 a.m.)
Deposited	1 year, 10 months ago (Oct. 12, 2023, 3:45 p.m.)
Indexed	1 year ago (Aug. 6, 2024, 12:39 a.m.)
Issued	20 years, 11 months ago (Sept. 15, 2004)
Published	20 years, 11 months ago (Sept. 15, 2004)
Published Online	20 years, 11 months ago (Sept. 15, 2004)
Published Print	20 years, 10 months ago (Oct. 1, 2004)

Funders 0

None

BibTeX

@article{ZHANG_2004, title={Direct visualization of receptor arrays in frozen‐hydrated sections and plunge‐frozen specimens of E. coli engineered to overproduce the chemotaxis receptor Tsr}, volume={216}, ISSN={1365-2818}, url={http://dx.doi.org/10.1111/j.0022-2720.2004.01395.x}, DOI={10.1111/j.0022-2720.2004.01395.x}, number={1}, journal={Journal of Microscopy}, publisher={Wiley}, author={ZHANG, P. and BOS, E. and HEYMANN, J. and GNAEGI, H. and KESSEL, M. and PETERS, P. J. and SUBRAMANIAM, S.}, year={2004}, month=sep, pages={76–83} }

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