Golgi -mannosidases regulate cell surface N-glycan type and ectodomain shedding of the transmembrane protease corin

Corin is a transmembrane protease that activates natriuretic peptides on the cell membrane. Reduced cell surface targeting or increased ectodomain shedding disrupts cell membrane homeostasis of corin, thereby impairing its cell surface expression and enzyme activity. N-glycans are essential in corin ectodomain shedding. Lack of N-glycans promotes corin ectodomain shedding in the juxtamembrane and frizzled -1 domains. The nascent N-glycans, transferred onto the polypeptide of corin, undergo multistep N-glycan processing in the endoplasmic reticulum and Golgi. It remains unclear how trimming by Golgi alpha-mannosidases, the critical N-glycan processing steps in N-glycan maturation, may regulate corin biosynthesis. In this study, we examined the effects of kifunensine and swainsonine, the inhibitors for alpha-mannosidases I and II, on corin expression and function. Western analysis of corin proteins in cell lysates and conditioned media from the inhibitor -treated corin-stable HEK293 cells and AC16 cells showed that both alpha-mannosidases I and II were required to maintain complex N-glycans on cell surface corin and protect corin from ectodomain shedding in the juxtamembrane and frizzled -1 domains. Cell viability analysis revealed that inhibition of alpha-mannosidase I or II sensitized cardiomyocytes to hydrogen peroxide -induced injury via regulating corin. Moreover, either one of the two coding genes was sufficient to perform Golgi alpha-mannosidase I trimming of N-glycans on corin. Similarly, this sufficiency was observed in Golgi alpha- mannosidase II -coding genes. Inhibition of ectodomain shedding restored corin zymogen activation from kifunensine- or swainsonine-induced reduction. Together, our results show the important roles of Golgi alpha-mannosidases in maintaining cell membrane homeostasis and biological activities of corin.