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Competing Pathways and Multiple Folding Nuclei in a Large Multidomain Protein, Luciferase

Biophys. J.. 2017-05; 
SchollZackary N, YangWeitao, MarszalekPio
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Mutagenesis Services … literature (47, 48). Truncated versions of Luciferase were generated at the DNA level through mutagenesis or gene synthesis from GenScript (Piscataway, NJ) at the residues specified in the main text. Plasmids were transformed … Get A Quote

摘要

Proteins obtain their final functional configuration through incremental folding with many intermediate steps in the folding pathway. If known, these intermediate steps could be valuable new targets for designing therapeutics and the sequence of events could elucidate the mechanism of refolding. However, determining these intermediate steps is hardly an easy feat, and has been elusive for most proteins, especially large, multidomain proteins. Here, we effectively map part of the folding pathway for the model large multidomain protein, Luciferase, by combining single-molecule force-spectroscopy experiments and coarse-grained simulation. Single-molecule refolding experiments reveal the initial nuc... More

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