Atomic force microscope of bacteriophage T4 and its tube‐baseplate complex

Ikai, Atsushi; Yoshimura, Kousei; Arisaka, Fumio; Ritani, Akishige; Imai, Kanae

doi:10.1016/0014-5793(93)81756-p

Cited by 26 publications

(14 citation statements)

References 14 publications

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“…Although this is not the first AFM investigation of T4, 40,41 this is the first detailed, high-resolution AFM imaging of bacteriophage T4 and its structural components. We have, in previous papers, presented AFM studies of a range of different viruses, 42 but none exhibiting the asymmetry and level of architectural sophistication associated with tailed bacteriophage.…”

Section: Discussionmentioning

confidence: 96%

Investigation of bacteriophage T4 by atomic force microscopy

2011

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Bacteriophage T4 was visualized using atomic force microscopy (AFM). The images were consistent with, and complementary to electron microscopy images. Head heights of dried particles containing DNA were about 75 nm in length and 60 nm in width, or about 100 nm and 85 nm respectively when scanned in fluid. The diameter of hydrated tail assemblies was 28 nm and their lengths about 130 nm. Seven to eight pronounced, right-handed helical turns with a pitch of 15 nm were evident on the tail assemblies. At the distal end of the tail was a knob shaped mass, presumably the baseplate. The opposite end, where the tail assembly joins the head, was tapered and connected to the portal complex, which was also visible. Phage that had ejected their DNA revealed the internal injection tube of the tail assembly. Heads disrupted by osmotic shock yielded boluses of closely packed DNA that unraveled slowly to expose threads composed of multiple twisted strands of nucleic acid. Assembly errors resulted in the appearance of several percent of the phage exhibiting two rather than one tail assemblies that were consistently oriented at about 72° to one another. No pattern of capsomeres was visible on native T4 heads. A mutant that is negative for the surface proteins hoc and soc, however, clearly revealed the icosahedral arrangement of ring shaped capsomeres on the surface. The hexameric rings have an outside diameter of about 14 nm, a pronounced central depression, and a center-to-center distance of 15 nm. Phage collapsed on cell surfaces appeared to be dissolving, possibly into the cell membrane.

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Section: Discussionmentioning

confidence: 96%

Investigation of bacteriophage T4 by atomic force microscopy

2011

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“…Atomic force microscope has been used to obtain high-resolution images of virus that have been immobilized on a variety of surfaces such as glass, mica, silicon, and Lang-muir-Blodgett films (Kuznetsov et al 2001;Ikai et al 1993). AFM permits discrimination among viruses based on their shape and size.…”

Section: Discussionmentioning

confidence: 99%

Integrity of a recombinant hemagglutinin protein of an avian influenza virus

Williams

Singh

et al. 2009

Biotechnol Lett

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An open reading frame representing cDNA from a hemagglutinin (HA) encoding gene of a low pathogenic avian influenza virus (AIV) subtype H10N7 was cloned in the pNMT1-TOPO vector under the control of thiamine response promoter. This construct was designated as pNMT1-HA. The pNMT1-HA construct was transformed into Schizosaccharomyces pombe for expression of HA antigen. The correct expression of recombinant HA protein was confirmed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. The level of expression of recombinant HA protein was approximately 0.2% of total soluble protein. Purified yeast-derived recombinant HA protein showed hemagglutination activity. The 2-D and 3-D scanning images of recombinant HA protein were observed with an atomic force microscope (AFM). The structural integrity of the HA protein under AFM and hemagglutination activity provided support that the recombinant HA protein may be suitable for development of AIV subunit vaccine for mass administration to poultry.

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“…A considerable number of investigators have therefore applied AFM to the observation of organic monolayers and bilayers Weisenhorn et al, 1990;Singh & Keller, 1991), ribonucleic and deoxyribonucleic acid (Thundat et al, 1992(Thundat et al, , 1993Hansma et al, 1992Hansma et al, , 1993bYang et al, 1992), amino acid crystals and proteins (Gould et al, 1988;Drake et al, 1989;Arakawa et al, 1992). There are also several reports on AFM observation of viruses, phages and in vitro cells (Butt et al, 1990;Gould et al, 1990;Henderson et al, 1992;Ikai et al, 1993;Kasas et al, 1993;Parpura et al, 1993). However, AFM studies on cells and tissues in situ have been limited in number, owing to technical difficulties in preparing adequate specimens.…”

Section: Discussionmentioning

confidence: 99%

X‐ray microscopy and imaging of Escherichia coli, LPS and DNA

Rajyaguru

Kado

Torres

et al. 1997

Journal of Microscopy

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SummaryUltrastructural examination by transmission and scanning electron microscopy involves a series of specialized preparation steps which may introduce artefacts in the micrographs. X-ray microscopy can take instant images of specimens but is mostly restricted to a few synchrotron X-ray sources. We have utilized a bench-top nanosecond laserplasma to produce a single-shot source of nanosecond X-rays tuned for maximum contrast with carbon-rich material. To examine the ultrastructure by absorption profiles, we utilized a laser-produced plasma generated by a single-shot laser (1 . 06 mm wavelength, 5 × 10 12 W cm ¹2 intensity) focused on to a silicon target as an X-ray source for high-resolution X-ray microscopy. This approach eliminates the specimen preparation steps. Whole hydrated cells of Escherichia coli and purified preparations of lipopolysaccharide (LPS) and chromosomal DNA (cDNA) were streaked onto poly(methyl methacrylate) (PMMA)-coated grids (resist). This resist was exposed to X-rays under vacuum at a distance of 2 . 5 cm from the target disc. The silicon plasma produced by a 10-ns burst of laser energy (at 20 J) radiates strong emission lines in the region of 300 eV. The X-rays penetrate the sample and their absorption profile is transferred on to the resist where PMMA acts as a negative to generate an image. By atomic force microscopy imaging of this photoresist we have visualized layers around cells of E. coli, darker areas inside the cell probably corresponding to cDNA, and preliminary images of LPS and DNA molecules. This technique has resolution at the 100 Å level, produces images similar to the space-filling models of macromolecules and may be of great value in the study of the ultrastructure of hydrated live biological specimens.

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Atomic force microscope of bacteriophage T4 and its tube‐baseplate complex

Cited by 26 publications

References 14 publications

Investigation of bacteriophage T4 by atomic force microscopy

Investigation of bacteriophage T4 by atomic force microscopy

Integrity of a recombinant hemagglutinin protein of an avian influenza virus

X‐ray microscopy and imaging of Escherichia coli, LPS and DNA

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