The complex folding network of single calmodulin molecules |
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Authors: | Stigler Johannes Ziegler Fabian Gieseke Anja Gebhardt J Christof M Rief Matthias |
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Institution: | Physik Department E22, Technische Universit?t München, James-Franck-Strasse, 85748 Garching, Germany. |
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Abstract: | Direct observation of the detailed conformational fluctuations of a single protein molecule en route to its folded state has so far been realized only in silico. We have used single-molecule force spectroscopy to study the folding transitions of single calmodulin molecules. High-resolution optical tweezers assays in combination with hidden Markov analysis reveal a complex network of on- and off-pathway intermediates. Cooperative and anticooperative interactions across domain boundaries can be observed directly. The folding network involves four intermediates. Two off-pathway intermediates exhibit non-native interdomain interactions and compete with the ultrafast productive folding pathway. |
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