Two engineered eglin c mutants potently and selectively inhibiting kexin or furin
Basic science published in FEBS Lett (2004)
Abstract
Eglin c with mutants L45R and D42R at the P(1) and P(4) positions has been reported to become a stable inhibitor toward the proprotein convertases (PC), furin and kexin, with a K(i) of 2.3x10(-8) and 1.3x10(-10) M, respectively. The mutant was further engineered at the P(2)'-P(4)' positions to create a more potent and selective inhibitor for each enzyme. The residue Asp at P(1)' which is crucial for stabilizing the conformation of eglin c remained unchanged. The eglin c mutants cloned into the vector pGEX-2T and expressed in Escherichia coli (DH5alpha) were purified to homogeneity, and their inhibitory activities toward the purified recombinant furin and kexin were examined. The results showed that (1) Leu47 at P(2)' replaced with either a positively or negatively charged residue resulted in a decrease in inhibitory activities to both enzymes; (2) the replacement of Arg with Asp at P(3)' was favorable for inhibiting furin with a K(i) of 7.8 x 10(-9) M, but not for inhibiting kexin; (3) the replacement of Tyr with Glu at P(4)' increased the inhibitory activity to kexin with a K(i) of 3 x 10(-11) M, but was almost without any influence on furin inhibition. It was indicated that the inhibitory specificity of eglin c could be changed from inhibiting elastase to inhibiting PCs by site-directed mutation at the P positions, while the inhibitory selectivity to furin or kexin could be optimized by mutation at the P' positions.
Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.
Resumen
Site-directed mutagenesis of eglin C yields potent furin (Ki 7.8x10^-9 M) and kexin (Ki 3x10^-11 M) inhibitors.
Por qué esto importa para la hirudoterapia
Este estudio examinó cómo las mutaciones dirigidas a sitio en las posiciones P′ de la eglina c podrían optimizar la selectividad inhibitoria para las proproteínas convertasas furina y kexina, reportando mutantes diseñados con valores de Ki de 7.8×10⁻⁹ M para furina y 3×10⁻¹¹ M para kexina, y demostrando que la especificidad inhibitoria podría desplazarse desde la elastasa hacia las proproteínas convertasas. La eglina c es un inhibidor de serina proteasa aislado originalmente de la sanguijuela medicinal Hirudo medicinalis, por lo que este trabajo de ingeniería de proteínas es directamente relevante para el interés de ASH en caracterizar y readaptar terapéuticamente los componentes bioactivos del secretoma de la sanguijuela. El rediseño racional de un inhibidor derivado de la sanguijuela para dirigirse a proteasas específicas relevantes para enfermedades puede informar el desarrollo de biofármacos dirigidos que se basen en moléculas derivadas de la hirudoterapia. Sin embargo, este es un estudio bioquímico in vitro que utiliza enzimas purificadas recombinantes e inhibidores expresados en E. coli, no un estudio clínico ni de eficacia in vivo, por lo que no establece la utilidad terapéutica en la terapia con sanguijuelas en sí misma.
Citación
Two engineered eglin c mutants potently and selectively inhibiting kexin or furin.
Liu Z et al. · FEBS letters, 2004
Contexto clínico relacionado
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Añadido a la biblioteca ASH: May 27, 2026 · Última actualización del sitio: 18 de junio de 2026