Direct discrimination between models of protein activation by single-molecule force measurements |
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| Authors: | Nevo Reinat Brumfeld Vlad Elbaum Michael Hinterdorfer Peter Reich Ziv |
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| Affiliation: | Department of Biological Chemistry, Weizmann Institute of Science, Rehovot, Israel. |
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| Abstract: | The limitations imposed on the analyses of complex chemical and biological systems by ensemble averaging can be overcome by single-molecule experiments. Here, we used a single-molecule technique to discriminate between two generally accepted mechanisms of a key biological process--the activation of proteins by molecular effectors. The two mechanisms, namely induced-fit and population-shift, are normally difficult to discriminate by ensemble approaches. As a model, we focused on the interaction between the nuclear transport effector, RanBP1, and two related complexes consisting of the nuclear import receptor, importin beta, and the GDP- or GppNHp-bound forms of the small GTPase, Ran. We found that recognition by the effector proceeds through either an induced-fit or a population-shift mechanism, depending on the substrate, and that the two mechanisms can be differentiated by the data. |
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