Publikation: Conformational Plasticity and Dynamics in the Generic Protein Folding Catalyst SlyD Unraveled by Single-Molecule FRET
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The relation between conformational dynamics and chemistry in enzyme catalysis recently has received increasing attention. While, in the past, the mechanochemical coupling was mainly attributed to molecular motors, nowadays, it seems that this linkage is far more general. Single-molecule fluorescence methods are perfectly suited to directly evidence conformational flexibility and dynamics. By labeling the enzyme SlyD, a member of peptidyl-prolyl cis-trans isomerases of the FK506 binding protein type with an inserted chaperone domain, with donor and acceptor fluorophores for single-molecule fluorescence resonance energy transfer, we directly monitor conformational flexibility and conformational dynamics between the chaperone domain and the FK506 binding protein domain. We find a broad distribution of distances between the labels with two main maxima, which we attribute to an open conformation and to a closed conformation of the enzyme. Correlation analysis demonstrates that the conformations exchange on a rate in the 100 Hz range. With the aid from Monte Carlo simulations, we show that there must be conformational flexibility beyond the two main conformational states. Interestingly, neither the conformational distribution nor the dynamics is significantly altered upon binding of substrates or other known binding partners. Based on these experimental findings, we propose a model where the conformational dynamics is used to search the conformation enabling the chemical step, which also explains the remarkable substrate promiscuity connected with a high efficiency of this class of peptidyl-prolyl cis-trans isomerases.
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KAHRA, Dana, Michael KOVERMANN, Christian LÖW, Verena HIRSCHFELD, Caroline HAUPT, Jochen BALBACH, Christian Gerhard HÜBNER, 2011. Conformational Plasticity and Dynamics in the Generic Protein Folding Catalyst SlyD Unraveled by Single-Molecule FRET. In: Journal of Molecular Biology (JMB). 2011, 411(4), pp. 781-790. ISSN 0022-2836. eISSN 1089-8638. Available under: doi: 10.1016/j.jmb.2011.05.002BibTex
@article{Kahra2011-08-26Confo-44626, year={2011}, doi={10.1016/j.jmb.2011.05.002}, title={Conformational Plasticity and Dynamics in the Generic Protein Folding Catalyst SlyD Unraveled by Single-Molecule FRET}, number={4}, volume={411}, issn={0022-2836}, journal={Journal of Molecular Biology (JMB)}, pages={781--790}, author={Kahra, Dana and Kovermann, Michael and Löw, Christian and Hirschfeld, Verena and Haupt, Caroline and Balbach, Jochen and Hübner, Christian Gerhard} }
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