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

Journal of Microscopy
Peijun ZhangSriram Subramaniam

Abstract

We 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 degrees C using a diamond knife with a 25 degrees 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.

Citations

Jun 21, 2008·Histochemistry and Cell Biology·Vladan LucićWolfgang Baumeister
Aug 4, 2009·Histochemistry and Cell Biology·Jason PiersonPeter J Peters
Jun 21, 2005·Journal of Structural Biology·Giovanni CardoneAlasdair C Steven
Jun 9, 2005·Current Opinion in Microbiology·Sriram Subramaniam
Nov 16, 2011·Quarterly Reviews of Biophysics·Lu Gan, Grant J Jensen
Feb 17, 2006·Nature Reviews. Molecular Cell Biology·Stephan NickellWolfgang Baumeister
Aug 12, 2009·Nature Reviews. Microbiology·Jacqueline L S Milne, Sriram Subramaniam
Dec 16, 2005·Proceedings of the National Academy of Sciences of the United States of America·Benoît ZuberJacques Dubochet
Mar 16, 2007·Proceedings of the National Academy of Sciences of the United States of America·Peijun ZhangSriram Subramaniam
Mar 5, 2008·Proceedings of the National Academy of Sciences of the United States of America·Christian HoffmannHarald Engelhardt
Oct 7, 2009·Proceedings of the National Academy of Sciences of the United States of America·Ariane BriegelGrant J Jensen
Aug 24, 2011·Journal of Electron Microscopy·Jason PiersonPeter J Peters
Jun 3, 2010·Cold Spring Harbor Perspectives in Biology·Elitza I TochevaGrant J Jensen
Mar 18, 2008·Journal of Bacteriology·Gerhard WannerJörg Overmann
Jan 22, 2008·Journal of Bacteriology·Robert BelasZhaomin Yang
Jul 22, 2006·Molecular Microbiology·David KentnerVictor Sourjik
Jan 6, 2007·Journal of Microscopy·Mark S LadinskyJ Richard McIntosh
Feb 6, 2008·Molecular Microbiology·Gavin E MurphyGrant J Jensen
Oct 7, 2004·Current Opinion in Neurobiology·Richard D Leapman
Nov 9, 2006·Ultrastructural Pathology·M Joseph Costello
Mar 1, 2007·Methods in Cell Biology·Jacques DubochetFrançoise Livolant
May 18, 2005·Biochimica Et Biophysica Acta·Brad J Marsh
Sep 25, 2007·Journal of Structural Biology·Harald Engelhardt
Aug 24, 2006·BioTechniques·Kent L McDonald, Manfred Auer
Mar 28, 2008·Molecular Microbiology·Ariane BriegelGrant J Jensen
Nov 28, 2012·The FEBS Journal·Jacqueline L S MilneSriram Subramaniam
Apr 4, 2021·Biomolecules·Alister BurtIrina Gutsche
Oct 24, 2007·Langmuir : the ACS Journal of Surfaces and Colloids·Laurent SagalowiczMartin Michel
Feb 9, 2007·Langmuir : the ACS Journal of Surfaces and Colloids·David J MontefuscoRobert M Weis

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.