Abstract
The study of protein folding has been conventionally hampered by the assumption that all single-domain proteins fold by an all-or-none process (two-state folding) that makes it impossible to resolve folding mechanisms experimentally. Here we describe an experimental method for the thermodynamic analysis of protein folding at atomic resolution using nuclear magnetic resonance (NMR). The method is specifically developed for the study of small proteins that fold autonomously into basic supersecondary structure motifs, and that do so in the sub-millisecond timescale (folding archetypes). From the NMR experiments we obtain hundreds of atomic unfolding curves that are subsequently analyzed leading to the determination of the characteristic network of folding interactions. The application of this approach to a comprehensive catalog of elementary folding archetypes holds the promise of becoming the first experimental approach capable of unraveling the basic rules connecting protein structure and folding mechanism.
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Acknowledgments
This work was supported by the Marie Curie Excellence Award MEXT-CT-2006-042334, and the grants BFU2008-03237, BFU2008-03278 and CONSOLIDER CSD2009-00088 from the Spanish Ministry of Science and Innovation (MICINN).
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Sborgi, L., Verma, A., Sadqi, M., de Alba, E., Muñoz, V. (2012). Protein Folding at Atomic Resolution: Analysis of Autonomously Folding Supersecondary Structure Motifs by Nuclear Magnetic Resonance. In: Kister, A. (eds) Protein Supersecondary Structures. Methods in Molecular Biology, vol 932. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-065-6_13
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DOI: https://doi.org/10.1007/978-1-62703-065-6_13
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