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Please use this identifier to cite or link to this item: http://hdl.handle.net/1959.3/216079
- Single molecule folding of RNase H on biocompatible surfaces
- Amirgoulova, Elza; Heyes, Colin D.; Groll, Juergen; Kobitski, Andrei Yu; Ameringer, Thomas; Roecker, Carlheinz; Moeller, Martin; Nienhaus, G. Ulrich
- The heterogeneity of individual trajectories of a protein adopting its native conformation from the myriad of denatured states can be studied by single molecule spectroscopy. However, to study time-scales longer than a few milliseconds, the protein has to be fixed in space, such as immobilization onto a surface. One major problem in immobilization is that the surface must not interfere with the protein, nor introduce additional heterogeneities. Here we characterize and compare several surfaces designed for single molecule experiments for biocompatibility. We examine surface homogeneity, their resistance to non-specific protein adsorption and their effect on protein conformation. Using single molecule FRET, we also investigate the ability of the immobilized protein to fold back into its native structure after denaturation. It is equally important that the surface does not interfere with the energetics of this folding process. Therefore, we investigate this by following various thermodynamic and kinetic parameters of the immobilized single proteins during the unfolding-refolding process. We discuss such surfaces that fulfill these requirements for single RNase H molecules, a model protein used in folding studies.
- Publication type
- Journal article
- Biophysical Journal: incorporating abstracts from the 48th Annual Meeting of the Biophysical Society, Baltimore, Maryland, United States, 14-18 February 2004, Vol. 86, supplement 1 (Jan 2004), p. 473A
- Publication year
- FOR Code(s)
- 02 Physical Sciences; 03 Chemical Sciences; 06 Biological Sciences
- Biocompatibility; Protein conformation
- Cell Press
- Publisher URL
- Copyright © 2004.
- Peer reviewed