Protein unfolding: Denaturant vs. force

Colleen Kelly, Matthew J. Gage

Research output: Contribution to journalArticlepeer-review

Abstract

While protein refolding has been studied for over 50 years since the pioneering work of Christian Anfinsen, there have been a limited number of studies correlating results between chemical, thermal, and mechanical unfolding. The limited knowledge of the relationship between these processes makes it challenging to compare results between studies if different refolding methods were applied. Our current work compares the energetic barriers and folding rates derived from chemical, thermal, and mechanical experiments using an immunoglobulin-like domain from the muscle protein titin as a model system. This domain, I83, has high solubility and low stability relative to other Ig domains in titin, though its stability can be modulated by calcium. Our experiments demonstrated that the free energy of refolding was equivalent with all three techniques, but the refolding rates exhibited differences, with mechanical refolding having slightly faster rates. This suggests that results from equilibrium-based measurements can be compared directly but care should be given comparing refolding kinetics derived from refolding experiments that used different unfolding methods.

Original languageEnglish (US)
Article number1395
JournalBiomedicines
Volume9
Issue number10
DOIs
StatePublished - Oct 2021
Externally publishedYes

Keywords

  • Chemical denaturation
  • Immunoglobulin domain
  • Magnetic tweezers
  • Protein refolding
  • Thermal denaturation
  • Titin

ASJC Scopus subject areas

  • Medicine (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)

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