Design of an optical switch for studying conformational dynamics in individual molecules of GroEL |
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Authors: | Frank Gabriel A Kipnis Yakov Smolensky Elena Daube Shirley S Horovitz Amnon Haran Gilad |
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Institution: | Department of Chemical Physics, Weizmann Institute of Science, Rehovot 76100, Israel. |
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Abstract: | We describe the design of an optical switch in the chaperonin GroEL that is opened and closed by its ATP- and cochaperonin GroES-driven conformational changes. The switch, based on a fluorophore and a quencher, is engineered into the single-ring variant of the chaperone, and shows dramatic modulation of its fluorescent intensity in response to the transition of the protein between its allosteric states. It, therefore, forms a sensitive probe for the dynamics of the allosteric transitions of this machine, both in the bulk and in single molecules. |
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