Comparative kinetic characterization of the activity of glycosylated and non-glycosylated trypsin-like serine protease isolated from adults of Rhyzopertha dominica (Coleoptera: Bostrichidae) reared on the grain of three diff erent cultivars of wheat

Fernanda S. Zavala-Ibarra, Aldo A. Arvizu-Flores*, Oliviert Martínez-Cruz, Pablo S. Osuna-Amarillas, Jose L. Cárdenas-López, Carmen L. Del-Toro-Sánchez, Carlos R. González-Ruiz, Jose A. Tapia-Hernández, Rey D. Iturralde-García, Francisco J. Cinco-Moroyoqui*

*Autor correspondiente de este trabajo

Producción científica: Contribución a una revistaArtículorevisión exhaustiva

Resumen

Rhyzopertha dominica is a pest that uses trypsin-like serine protease enzymes to hydrolyse the proteins in the cereal grains on which it feeds. The present study reveals for the fi rst time that that there are both glycosylated and non-glycosylated serine proteases. The progeny of R. dominica reared on the grain three varieties of wheat were used to fractionate their trypsinlike serine proteases using Concanavalin A affi nity chromatography. The albumin fractions from the wheat cultivars used in this study were subjected to size exclusion chromatography to fractionate the albumin inhibitors that are highly specifi c for the serine protease activity of R. dominica. Kinetic and thermodynamic assays were used to diff erentiate both types of enzymes. In general, the catalytic effi ciency values Vmax/Km for glycosylated proteases were higher, indicating that glycosylation increases the affi nity for the substrate. Inhibition assays using wheat albumins revealed that the glycosylated enzymes had higher Ki values, indicating a low affi nity for the inhibitors than the non-glycosylated enzymes. Thermodynamic analysis indicates that glycosylation increases the activation energy Ea improving the serine proteases’ catalysis. Thus it is likely that R. dominica uses glycosylated proteases in order to optimize the hydrolysis of cereal proteins and nullify the action of wheat grain protease inhibitors and increase its chances of survival.

Idioma originalInglés
Páginas (desde-hasta)233-243
Número de páginas11
PublicaciónEuropean Journal of Entomology
Volumen120
DOI
EstadoPublicada - 2023

Nota bibliográfica

Publisher Copyright:
© Institute of Entomology, Biology Centre, Czech Academy of Sciences, České Budějovice. An Open Access article distributed under the Creative Commons (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).

Huella

Profundice en los temas de investigación de 'Comparative kinetic characterization of the activity of glycosylated and non-glycosylated trypsin-like serine protease isolated from adults of Rhyzopertha dominica (Coleoptera: Bostrichidae) reared on the grain of three diff erent cultivars of wheat'. En conjunto forman una huella única.

Citar esto