Improvement of the specificity of dipetarudin by site directed mutagenesis
Research article published in Thrombosis and haemostasis (2005)
Abstract
Protease specificity is crucial to the design of thrombin inhibitors as inhibition of other physiologically relevant serine-proteases can compromise their clinical use. Dipetarudin, a potent thrombin inhibitor, also inhibits trypsin and plasmin. Due to the specificity of an inhibitor being influenced by the amino acid residue at the P1 position, we replaced the Arg10 at P1 position of dipetarudin by a histidine, which is the P1 residue of rhodniin, a very specific thrombin inhibitor. The amino acid replacement was carried out by site directed mutagenesis. The mutant, dipetarudinR10H, showed a loss of plasmin and trypsin inhibitory activities present in its wild-type counterpart and a 3-fold higher dissociation constant for thrombin than dipetarudin. However, compared to dipetarudin and r-hirudin, dipetarudinR10H showed similar activity in coagulation screening assays such as activated partial thromboplastin time (aPTT), prothrombin time (PT), ecarin clotting time (ECT) and ecarin chromogenic assay (ECA).
Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.
Resumen
Protease specificity is crucial to the design of thrombin inhibitors as inhibition of other physiologically relevant serine-proteases can compromise their clinical use.
Por qué esto importa para la hirudoterapia
Este estudio utilizó mutagénesis dirigida al sitio para sustituir la arginina P1 en la posición 10 de la dipetarudina por histidina, generando un mutante (dipetarudinaR10H) que perdió la actividad inhibidora frente a la plasmina y la tripsina. En ensayos de cribado de coagulación, incluidos el tiempo de tromboplastina parcial activada y el tiempo de protrombina, la dipetarudinaR10H mostró una actividad similar tanto a la de la dipetarudina como a la de la r-hirudina. El trabajo es relevante porque evalúa inhibidores de la trombina obtenidos por ingeniería genética y compara directamente sus efectos funcionales sobre la coagulación frente a la r-hirudina. Sin embargo, el resumen no identifica el origen biológico de la r-hirudina ni de la dipetarudina, ni involucra sanguijuelas vivas. Por tanto, la conexión con el secretoma de la sanguijuela es totalmente indirecta, dependiendo exclusivamente de la comparación in vitro con la r-hirudina.
Citación
Improvement of the specificity of dipetarudin by site directed mutagenesis
Lpez M et al. · Thrombosis and haemostasis, 2005
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Añadido a la biblioteca ASH: May 27, 2026 · Última actualización del sitio: 18 de junio de 2026