Diselenide crosslinks for enhanced and simplified oxidative protein folding
Protein chemistry study published in Communications Chemistry (2021)
Abstract
The in vitro oxidative folding of proteins has been studied for over sixty years, providing critical insight into protein folding mechanisms. Hirudin, the most potent natural inhibitor of thrombin, is a 65-residue protein with three disulfide bonds, and is viewed as a folding model for a wide range of disulfide-rich proteins. Hirudin's folding pathway is notorious for its highly heterogeneous intermediates and scrambled isomers, limiting its folding rate and yield in vitro. Aiming to overcome these limitations, we undertake systematic investigation of diselenide bridges at native and non-native positions and investigate their effect on hirudin's folding, structure and activity. Our studies demonstrate that, regardless of the specific positions of these substitutions, the diselenide crosslinks enhanced the folding rate and yield of the corresponding hirudin analogues, while reducing the complexity and heterogeneity of the process. Moreover, crystal structure analysis confirms that the diselenide substitutions maintained the overall three-dimensional structure of the protein and left its function virtually unchanged. The choice of hirudin as a study model has implications beyond its specific folding mechanism, demonstrating the high potential of diselenide substitutions in the design, preparation and characterization of disulfide-rich proteins.
Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.
Summary
Diselenide-crosslink substitutions in hirudin enhance folding rate and yield while reducing folding-intermediate heterogeneity — important chemistry for recombinant hirudin manufacturing.
Why This Matters for Hirudotherapy
This in vitro study investigated the oxidative folding pathways of the protein hirudin — described in the abstract as the most potent natural inhibitor of thrombin and a 65-residue protein with three disulfide bonds — by substituting diselenide bridges at native and non-native positions. The substitutions enhanced the folding rate and yield of hirudin analogues while maintaining the three-dimensional structure and leaving function virtually unchanged. The relevance to ASH's domain is weak and indirect: although hirudin is widely known as a leech-derived protein, the abstract itself does not mention leeches, leech saliva, or hirudotherapy, and the study is purely focused on in vitro protein chemistry and folding mechanisms. The work provides no direct therapeutic application or clinical data regarding leech therapy.
Citation
Diselenide crosslinks for enhanced and simplified oxidative protein folding.
Mousa R et al. · Communications chemistry, 2021
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