Multi-Probe Investigation of Proteomic Structure of Pathogens

Malkin, Alexander J.; Plomp, Marco; Leighton, Terrance; Vogelstein, Bert; Wheeler, Katherine E.

doi:10.2172/926009

Cited by 1 publication

(3 citation statements)

References 9 publications

(4 reference statements)

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“…The first AFM studies of noncrystalline viruses were retroviruses on cell surfaces (20)(21)(22), again, principally because they were immobilized by their association with cell surfaces. Single particles of larger viruses, and helical viruses, were eventually visualized by AFM, and these included tobacco mosaic virus, cauliflower mosaic virus, Tipula iridescent virus (15,23), herpes simplex virus (24), vaccinia virus (25,26), and mimivirus (9). Although virus crystals were investigated using both contact and tapping mode, noncrystalline specimens were imaged exclusively with tapping mode, in both air and buffer.…”

Section: Afm Instrument Setupmentioning

confidence: 99%

“…As has been shown in AFM investigations of a number of viruses, it is, in fact, an invaluable tool for deducing the interior architecture of virions, regardless of their external form or size. This is because it is possible to strip away layers of structure systematically by chemical, physical, and enzymatic means (23,26,27) and to accompany this process of dissection by AFM visualization. Using the same strategy as that used by conventional anatomists, it has been proven possible to disassemble viral specimens, see what is inside, and ascertain how the components are linked.…”

Section: Virus Imagingmentioning

confidence: 99%

“…This is not true, however, as we can apply the same technique that has been used by anatomists for centuriesdissection. With the aid of chemical, enzymatic, and physical tools, we can systematically pare a complex entity, including a virus, down to its core, layer by layer (23). At each stage, AFM may then be used to visualize what remains and what has been removed as well (see Note 9).…”

Section: Topography As a Function Of Composition And Architecturementioning

confidence: 99%

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Atomic Force Microscopy Investigation of Viruses

McPherson¹,

Kuznetsov²

2011

Methods in Molecular Biology

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Atomic force microscopy (AFM) has proven to be a valuable approach to delineate the architectures and detailed structural features of a wide variety of viruses. These have ranged from small plant satellite viruses of only 17 nm to the giant mimivirus of 750 nm diameter, and they have included diverse morphologies such as those represented by HIV, icosahedral particles, vaccinia, and bacteriophages. Because it is a surface technique, it provides images and information that are distinct from those obtained by electron microscopy, and in some cases, at even higher resolution. By enzymatic and chemical dissection of virions, internal structures can be revealed, as well as DNA and RNA. The method is relatively rapid and can be carried out on both fixed and unfixed samples in either air or fluids, including culture media. It is nondestructive and even non-perturbing. It can be applied to individual isolated virus, as well as to infected cells. AFM is still in its early development and holds great promise for further investigation of biological systems at the nanometer scale.Key words: Imaging, Nanoscale, Structure, Infection, Nucleic acids, Icosahedra A direct imaging technology that promises to have a significant impact on structural biology, and which is, in most ways, complementary to X-ray diffraction and electron microscopy, the classical approaches, is atomic force microscopy (AFM) (1-3). An immediate advantage of AFM is that it is based on relatively simple physical principles, unlike X-ray crystallography, and the instruments are mechanically and electronically rather straightforward, unlike electron microscopy. Unlike both of the other technologies, AFM is fairly inexpensive to institute and apply, even to biological specimens. The acuity and investigative size range of the AFM have proven to be quite remarkable and it is now permitting researchers new access to virus structure and the effects of viruses on organisms. Indeed, it has allowed us to

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