Single-particle virology
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Single-particle virology Bálint Kiss 1 & Dorottya Mudra 1 Miklós Kellermayer 1
& György Török
1
& Zsolt Mártonfalvi
1
& Gabriella Csík
1
& Levente Herényi
1
&
Received: 7 July 2020 / Accepted: 18 August 2020 # The Author(s) 2020
Abstract The development of advanced experimental methodologies, such as optical tweezers, scanning-probe and super-resolved optical microscopies, has led to the evolution of single-molecule biophysics, a field of science that allows direct access to the mechanistic detail of biomolecular structure and function. The extension of single-molecule methods to the investigation of particles such as viruses permits unprecedented insights into the behavior of supramolecular assemblies. Here we address the scope of viral exploration at the level of individual particles. In an era of increased awareness towards virology, single-particle approaches are expected to facilitate the in-depth understanding, and hence combating, of viral diseases. Keywords Single-molecule mechanics . Atomic force microscopy . Optical tweezers . Super-resolution microscopy . Viral genome packaging . Viral genome release
Single-molecule and single-particle science Much of our knowledge in natural sciences is derived from ensembles of particles (atoms, molecules), the standard quantity of which is the mol (Van Holde et al. 1998). The properties and the behavior of the individual particles are thus extrapolations from ensemble average parameters. By contrast, single-molecule and single-particle science focuses on the individual (Bustamante et al. 2000; Kellermayer 2005). Hence, not only the average but also the distribution of the measured parameter can be obtained, which provides a direct insight into the structure, function, and dynamics of the investigated molecule or particle and into the mechanisms behind the processes the molecule or the particle is involved in. Investigation of molecules and particles one by one has particular significance in biological systems, considering that in a living cell often there are only a handful of molecules of the same species present. Although particles, in the biological sense, are usually supramolecular assemblies and therefore are composed of a number of molecules, similar methodological principles can be applied to them as to single molecules.
* Miklós Kellermayer [email protected] 1
Department of Biophysics and Radiation Biology, Semmelweis University, Budapest, Hungary
There are at least four areas in which single-molecule or single-particle techniques provide unique gain over ensemble methods. First, individuals can be identified in a crowd and followed in space and time. Considering the unusually dense and crowded environment of the intracellular space, singlemolecule and single-particle visualization methods must be applied to uncover the behavior of individual molecular species. Second, the temporal distribution of molecular states may be described; hence stochastic processes, such as the blinking of fluorescent proteins (Dickson e
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