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.
Resumen
Diselenide-crosslink substitutions in hirudin enhance folding rate and yield while reducing folding-intermediate heterogeneity — important chemistry for recombinant hirudin manufacturing.
Por qué esto importa para la hirudoterapia
Este estudio in vitro investigó las vías de plegamiento oxidativo de la proteína hirudina —descrita en el resumen como el inhibidor natural más potente de la trombina y una proteína de 65 residuos con tres puentes disulfuro— mediante la sustitución de puentes de diseleniuro en posiciones nativas y no nativas. Las sustituciones aumentaron la velocidad de plegamiento y el rendimiento de los análogos de hirudina, al tiempo que mantuvieron la estructura tridimensional y dejaron la función prácticamente sin cambios. La relevancia para el ámbito de la ASH es débil e indirecta: aunque la hirudina es ampliamente conocida como una proteína derivada de la sanguijuela, el propio resumen no menciona sanguijuelas, saliva de sanguijuela ni hirudoterapia, y el estudio se centra exclusivamente en la química de proteínas in vitro y en los mecanismos de plegamiento. El trabajo no aporta ninguna aplicación terapéutica directa ni datos clínicos relativos a la terapia con sanguijuelas.
Citación
Diselenide crosslinks for enhanced and simplified oxidative protein folding.
Mousa R et al. · Communications chemistry, 2021
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Añadido a la biblioteca ASH: May 27, 2026 · Última actualización del sitio: 18 de junio de 2026