Rational design of stable and functional hirudin III mutants with lower antigenicity
Research article published in Biologicals : journal of the International Association of Biological Standardization (2015)
Abstract
Hirudin is an inhibitor of thrombin and used as an effective anticoagulant, but has a potential to develop unacceptable immune responses. In this study, two computational tools were used to predict T-cell epitopes within Hirudin variant III (HVIII) sequence, and design mutations that would lessen its antigenicity. Homology models of native and mutant HVIII proteins (T4K, S9G, V21G, and V21K) were generated, and further used to assess their interactions with thrombin. The docking experiment showed that all mutants had a suitable pattern of interactions, with similar or lower interaction energies compared with the native protein. These complexes were subsequently subjected to molecular dynamics simulation. All mutants complexes had overall stable structures over simulation time, with RMSD, gyration radius, hydrogen bonds numbers, and accessible surface areas patterns that were comparable with the native HVIII over time. Interestingly, in all mutants, a shorter length was observed for the two salt bridges Arg73-Asp55 and Arg77-Glu57, which are suggested to be important in Hirudin-thrombin complex formation. Best selected mutants expressed in Escherichia coli BL21(DE3), subsequently SDS-PAGE and Western blot analysis confirmed the successful same expression of Hirudin and mutants. In conclusion, we believe that this computational approach could identify potentially safer proteins with preserved or even improved functionality.
Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.
Resumen
Hirudin is an inhibitor of thrombin and used as an effective anticoagulant, but has a potential to develop unacceptable immune responses.
Por qué esto importa para la hirudoterapia
Este estudio empleó herramientas computacionales (predicción de epítopos de linfocitos T, modelado por homología, acoplamiento molecular y simulación de dinámica molecular) para diseñar mutantes de la variante III de hirudina (T4K, S9G, V21G, V21K) destinados a reducir la antigenicidad preservando la función inhibidora de trombina. Los mutantes mostraron patrones adecuados de interacción con trombina y estructuras complejas estables, y los mutantes seleccionados se expresaron con éxito en E. coli BL21(DE3). Este trabajo es directamente relevante para ASH porque concierne a la ingeniería racional de la propia hirudina—el arquetípico anticoagulante de la sanguijuela—para mejorar su perfil de seguridad y terapéutico. Sin embargo, el estudio es enteramente computacional y de expresión preliminar in vitro, sin datos de bioensayo funcional, en animales ni clínicos; no se aborda su relevancia para la hirudoterapia viva.
Citación
Rational design of stable and functional hirudin III mutants with lower antigenicity
Asgari S et al. · Biologicals : journal of the International Association of Biological Standardization, 2015
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Añadido a la biblioteca ASH: May 27, 2026 · Última actualización del sitio: 18 de junio de 2026