A Single-Molecule Strategy to Capture Non-native Intramolecular and Intermolecular Protein Disulfide Bridges

Marc Mora*, Stephanie Board, Olivier Languin-Cattoën, Laura Masino, Guillaume Stirnemann, Sergi Garcia-Manyes

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

4 Citations (Scopus)

Abstract

Non-native disulfide bonds are dynamic covalent bridges that form post-translationally between two cysteines within the same protein (intramolecular) or with a neighboring protein (intermolecular), frequently due to changes in the cellular redox potential. The reversible formation of non-native disulfides is intimately linked to alterations in protein function; while they can provide a mechanism to protect against cysteine overoxidation, they are also involved in the early stages of protein multimerization, a hallmark of several protein aggregation diseases. Yet their identification using current protein chemistry technology remains challenging, mainly because of their fleeting reactivity. Here, we use single-molecule spectroscopy AFM and molecular dynamics simulations to capture both intra- and intermolecular disulfide bonds in γD-crystallin, a cysteine-rich, structural human lens protein involved in age-related eye cataracts. Our approach showcases the power of mechanical force as a conformational probe in dynamically evolving proteins and presents a platform to detect non-native disulfide bridges with single-molecule resolution.

Original languageEnglish
Pages (from-to)3922-3930
Number of pages9
JournalNano Letters
Volume22
Issue number10
DOIs
Publication statusPublished - 25 May 2022

Keywords

  • atomic force microscopy (AFM)
  • non-native disulfide bonds
  • protein folding
  • protein mechanochemistry
  • protein nanomechanics
  • single-molecule force spectroscopy

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