Temperature jump and fast photochemical oxidation probe submillisecond protein folding

Jiawei Chen, Don L. Rempel, Michael L. Gross

Research output: Contribution to journalArticlepeer-review

72 Scopus citations


We report a new mass-spectrometry-based approach for studying protein-folding dynamics on the submillisecond time scale. The strategy couples a temperature jump with fast photochemical oxidation of proteins (FPOP), whereby folding/unfolding is followed by changes in oxidative modifications by OH radical reactions. Using a flow system containing the protein barstar as a model, we altered the protein's equilibrium conformation by applying the temperature jump and demonstrated that its reactivity with OH free radicals serves as a reporter of the conformational change. Furthermore, we found that the time-dependent increase in mass resulting from free-radical oxidation is a measure of the rate constant for the transition from the unfolded to the first intermediate state. This advance offers the promise that, when extended with mass-spectrometry-based proteomic analysis, the sites and kinetics of folding/unfolding can also be followed on the submillisecond time scale.

Original languageEnglish
Pages (from-to)15502-15504
Number of pages3
JournalJournal of the American Chemical Society
Issue number44
StatePublished - Nov 10 2010


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