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Please use this identifier to cite or link to this item: http://hdl.handle.net/10261/60669
Title: Unequivocal single-molecule force spectroscopy of intrinsically disordered proteins
Authors: Oroz, Javier; Hervás, Rubén; Valbuena, Alejandro; Carrión-Vázquez, Mariano Sixto
Issue Date: 2012
Publisher: Humana Press
Citation: Methods in molecular biology (Clifton, N,J,) 896: 71- 87 (2012)
Abstract: Intrinsically disordered proteins (IDPs) are predicted to represent about one third of the eukaryotic proteome. The dynamic ensemble of conformations of this steadily growing class of proteins has remained hardly accessible for bulk biophysical techniques. However, single-molecule techniques provide a useful means of studying these proteins. Atomic force microscopy (AFM)-based single-molecule force spectroscopy (SMFS) is one of such techniques, which has certain peculiarities that make it an important methodology to analyze the biophysical properties of IDPs. However, several drawbacks inherent to this technique can complicate such analysis. We have developed a protein engineering strategy to overcome these drawbacks such that an unambiguous mechanical analysis of proteins, including IDPs, can be readily performed. Using this approach, we have recently characterized the rich conformational polymorphism of several IDPs. Here, we describe a simple protocol to perform the nanomechanical analysis of IDPs using this new strategy, a procedure that in principle can also be followed for the nanomechanical analysis of any protein. © 2012 Springer Science+Business Media New York.
URI: http://hdl.handle.net/10261/60669
DOI: 10.1007/978-1-4614-3704-8_5
Identifiers: doi: 10.1007/978-1-4614-3704-8_5
issn: 1064-3745
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