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Five challenges to bringing single-molecule force spectroscopy into living cells

Universite catholique de Louvain, Institute of Condensed Matter and Nanosciences, Louvain-la-Neuve, Belgium.
Nature Methods (Impact Factor: 25.95). 02/2011; 8(2):123-7. DOI: 10.1038/nmeth0211-123
Source: PubMed

ABSTRACT In recent years, single-molecule force spectroscopy techniques have been used to study how inter- and intramolecular interactions control the assembly and functional state of biomolecular machinery in vitro. Here we discuss the problems and challenges that need to be addressed to bring these technologies into living cells and to learn how cellular machinery is controlled in vivo.

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Available from: Daniel J Müller, Aug 15, 2015
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    • "Unauthenticated Download Date | 6/13/15 8:43 PM technique has been used to study how interand intramolecular interactions control the assembly and functional state of biomolecular machinery in vitro [34]. "
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    • "In this context, an AFM-based technique, single-molecule force spectroscopy (SMFS), has recently emerged in the field. In this technique, AFM tips interact with biomolecules immobilized on innate substrates or artificial biomembranes (in vitro studies) or present at the surface of living cells so to understand the intramolecular and intermolecular interactions of biomolecular systems (Müller et al., 2009; Dufrêne et al., 2011). SMFS techniques have been widely used in vitro, to monitor, for example, the interaction of cellular adhesion molecules, such as cadherins (Baumgartner et al., 2000) or oligosaccharides (Rief et al., 1997a) or to characterize the anchoring forces of peptides in lipid membranes (Ganchev et al., 2004). "
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    • "Recently, SMFS has been proven to be a useful method to analyze the activities of single molecules in situ on living bacteria [5] and mammalian cells [6]. However, current SMFS experiments are performed on purified biomolecules isolated from cell lines [7] or directly on cell lines [8]. Cell lines cultured in vitro are known to be quite different from cells in the human body, due to the huge difference between the in vitro and in vivo environment . "
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