Intermediates in membrane fusion and bilayer/nonbilayer phase transitions imaged by time-resolved cryo-transmission electron microscopy

Siegel, David P.; Burns, Janet L.; Chestnut, M. H.; Talmon, Yeshayahu

doi:10.1016/s0006-3495(89)82661-x

Cited by 150 publications

(105 citation statements)

References 38 publications

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“…2C (see also [23]). Dissociation of the globular HA1 subunits is'known to occur following low pH treatment' with many Influenza virus strains, including A/Aichi/2/68 [6].…”

Section: Proposalmentioning

confidence: 91%

An architecture for the fusion site of Influenza hemagglutinin

1990

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The recent finding that more than one InRuenza hemagglutinin (HA) is required at the fusion site for HA-expressing fibroblasts [l), together with the crystal structure of HA at neutral pH [2], provide the basic elements of a plausible model for this fusion site. Within an aggregate of HA trimers at low pH, we propose fusion intermediates which are based upon a minimal alteration to the known neutral pH structure of HA and which should have reasonable activation energies. This is the first model of a g~ycoprotein-media~d fusion site which explicitly accounts for the ~sposition of the lipids within these intermediates. While the fusion site created by HA will not be the same as that of eukaryotic fusion complexes [3], general characteristics could be shared.

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“…2C (see also [23]). Dissociation of the globular HA1 subunits is'known to occur following low pH treatment' with many Influenza virus strains, including A/Aichi/2/68 [6].…”

Section: Proposalmentioning

confidence: 91%

An architecture for the fusion site of Influenza hemagglutinin

1990

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“…Cryo‐TEM has been developed already in the seventies and eighties51, 52, 53 when it was realized that drying causes many different problems for soft materials and staining can introduce artefacts and is limited because its indirectness 54, 55. Other research fields that use cryo‐TEM are making more and more use of new technical developments such as the volta‐phase plate56 that enhances image contrast and allows imaging in focus.…”

Section: Quality and Outlookmentioning

confidence: 99%

Transmission Electron Microscopy as a Tool for the Characterization of Soft Materials: Application and Interpretation

2017

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Transmission electron microscopy (TEM) provides direct structural information on nano‐structured materials and is popular as a characterization tool in soft matter and supramolecular chemistry. However, technical aspects of sample preparation are overlooked and erroneous image interpretations are regularly encountered in the literature. There are three most commonly used TEM methods as we derived from literature: drying, staining and cryo‐TEM, which are explained here with respect to their application, limitations and interpretation. Since soft matter chemistry relies on a lot of indirect evidence, the role of TEM for the correct evaluation of the nature of an assembly is very large. Mistakes in application and interpretation can therefore have enormous impact on the quality of present and future studies. We provide helpful background information of these three techniques, the information that can and cannot be derived from them and provide assistance in selecting the right technique for soft matter imaging. This essay warns against the use of drying and explains why. In general cryo‐TEM is by far the best suited method and many mistakes and over‐interpretations can be avoided by the use of this technique.

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“…The interlamellar attachment-fusion hypothesis derives from the theoretical studies of Siegel (1986aSiegel ( , 1986bSiegel ( , 1986c and Siegel et al (1989). These studies suggest that there is a common structural intermediate in the formation of both the HIi phase and the interlamellar attachments, i.e.…”

Section: The Fracture-jump Lesionmentioning

confidence: 99%

A Contrast of the Plasma Membrane Lipid Composition of Oat and Rye Leaves in Relation to Freezing Tolerance

1994

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l h e lipid composition of the plasma membrane isolated from leaves of spring oat (Avena safiva 1. cv Ogle) was vastly different from that of winter rye (Secale cereale 1. cv Puma). l h e plasma membrane of spring oat contained large proportions of phospholipids (28.8 mol% of the total lipids), cerebrosides (27.2 mol%), and acylated sterylglucosides (27.3 mol%) with lesser proportions of free sterols (8.4 mol%) and sterylglucosides (5.6 mol%). In contrast, the plasma membrane of winter rye contained a greater proportion of phospholipids (36.6 mol%), and there was a lower proportion of cerebrosides (16.4 mol%); free sterols (38.1 mol%) were the predominant sterols, with lesser proportions of sterylglucosides (5.6 mol%) and acylated sterylglucosides (2.9 mol%). Although the relative proportions of individual phospholipids, primarily phosphatidylcholine and phosphatidylethanolamine, and the molecular species of these two phospholipids were similar in oat and rye, the relative proportions of di-unsaturated species of these two phospholipids were substantially lower in oat than in rye. l h e relative proportions of sterol species in oat were different from those in rye; the molecular species of cerebrosides were similar in oat and rye, with only slight differences in the proportions of the individual species. After 4 weeks of cold acclimation, the proportion of phospholipids increased significantly in both oat (from 28.8 to 36.8 mol%) and rye (from 36.6 to 43.3 mol%) as a result of increases in the proportions of phosphatidylcholine and phosphatidylethanolamine. For both oat and rye, the relative proportions of di-unsaturated species increased after cold acclimation, but the increase was greater in rye than in oat. In both oat and rye, this increase occurred largely during the first week of cold acclimation. During the 4 weeks of cold acclimation, there was a progressive decrease in the proportion of cerebrosides in the plasma membrane of rye (from 16.4 to 10.5 mol%), but there was only a small decrease in oat (from 27.2 to 24.2 mol%). In both oat and rye, there were only small changes in the proportions of free sterols and sterol derivatives during cold acclimation. Consequently, the proportions of both acylated sterylglucosides and cerebrosides remained substantially higher in oat than in rye after cold acclimation. l h e relationship between these differences in the plasma membrane lipid composition of oat and rye and their freezing tolerance is presented.

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Intermediates in membrane fusion and bilayer/nonbilayer phase transitions imaged by time-resolved cryo-transmission electron microscopy

Cited by 150 publications

References 38 publications

An architecture for the fusion site of Influenza hemagglutinin

An architecture for the fusion site of Influenza hemagglutinin

Transmission Electron Microscopy as a Tool for the Characterization of Soft Materials: Application and Interpretation

A Contrast of the Plasma Membrane Lipid Composition of Oat and Rye Leaves in Relation to Freezing Tolerance

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