A New Method for Rapid Characterization of the Folding Pathways of Multidisulfide-Containing Proteins

Ervin Welker, Laura Hathaway, Harold A. Scheraga

Research output: Contribution to journalArticle

13 Citations (Scopus)


Oxidative folding is a composite process that consists of both the conformational folding to the native three-dimensional structure and the regeneration of the native disulfide bonds of a protein, frequently involving over 100 disulfide intermediate species. Understanding the oxidative folding pathways of a multiple-disulfide-containing protein is a very difficult task that often requires years of devoted research due to the high complexity of the process and the very similar features of the large number of intermediates. Here we developed a method for rapidly delineating the major features of the oxidative folding pathways of a protein. The method examines the temperature dependence of the oxidative folding rate of the protein in combination with reduction pulses. Reduction pulses expose the presence of structured intermediates along the pathways. The correlation between the regeneration rate at different temperatures and the stability of the structured intermediates reveals the role that the intermediates play in determining the pathway. The method was first tested with bovine pancreatic ribonuclease A whose folding pathways were defined earlier. Then, it was explored to discern some of the major features of the folding pathways of its homologue, frog Onconase. The results suggest that the stability of the three-dimensional structure of the native protein is a major determinant of the folding rate in oxidative folding.

Original languageEnglish
Pages (from-to)3720-3721
Number of pages2
JournalJournal of the American Chemical Society
Issue number12
Publication statusPublished - Mar 31 2004

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Fingerprint Dive into the research topics of 'A New Method for Rapid Characterization of the Folding Pathways of Multidisulfide-Containing Proteins'. Together they form a unique fingerprint.

  • Cite this