The Francis Crick Institute
acs.nanolett.2c00043 (2).pdf (4.11 MB)

A single-molecule strategy to capture non-native intramolecular and intermolecular protein disulfide bridges.

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journal contribution
posted on 2022-05-27, 10:16 authored by Marc Mora, Stephanie Board, Olivier Languin-Cattoën, Laura Masino, Guillaume Stirnemann, Sergi Garcia-Manyes
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.


Crick (Grant ID: 10015, Grant title: STP Structural Biology)