Detection and characterization of a folding intermediate in barnase by NMR |
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Authors: | M Bycroft A Matouschek J T Kellis L Serrano A R Fersht |
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Institution: | Department of Chemistry, University of Cambridge, UK. |
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Abstract: | Protein engineering is being developed for mapping the energetics and pathway of protein folding. From kinetic studies on wild-type and mutant proteins, the sequence and energetics of formation of tertiary interactions of side chains can be mapped and the formation of secondary structure inferred. Here we cross-check and complement results from this approach by using a recently developed procedure that traps and characterizes secondary structure in intermediate states using 1H NMR. The refolding of barnase is shown to be a multiphasic process in which the secondary structure in alpha-helices and beta-sheets and some turns is formed more rapidly than is the overall folding. |
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