Key result
The loss of N-glycans in murine GASP-1 leads to a decrease in protein secretion rate but does not affect its ability to activate myogenesis in C2C12 cells.
Why the study?
Does N-glycosylation of murine GASP-1 affect its secretion and ability to activate myogenesis in C2C12 cells?
Does N-glycosylation of murine GASP-1 affect its secretion and ability to activate myogenesis in C2C12 cells?
N-glycosylation of murine GASP-1 is essential for its secretion rate but not for its biological activity in promoting myogenesis.
N-glycan loss impairs murine GASP-1 secretion without affecting myogenic activity in vitro; leaves open relevance to human muscle regeneration.
BACKGROUND/AIMS: Growth and differentiation factor-associated serum protein-1 (GASP-1) is a secreted protein known to be capable of binding and inhibiting the activity of several TGF-beta family members, including myostatin. The present study was designed to characterize murine GASP-1 post-translational modifications and to determine their influence on the biological activity of GASP-1. METHODS: We describe herein the site-directed mutagenesis of single N-glycosylation sites and combinations of them in 4 mutants of murine GASP-1. RESULTS: In vitro and in vivo analysis revealed that GASP-1 is a glycoprotein containing 2 N-glycans and several mucin-type O-glycans. Treatments by the recombinant murine GASP-1 protein enhance C2C12 proliferation and differentiation by inhibition of the myostatin pathway. The loss of N-glycans leads to a decrease in protein secretion rate but does not affect its ability to activate myogenesis. CONCLUSION: Analysis of structure-function relationships of murine GASP-1 provides insights into the involvement of the carbohydrate moiety of mGASP-1 on its biological activity.
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Brun et al. (2012) studied this question. Site-directed mutagenesis of N-glycosylation sites in murine GASP-1 was evaluated on Protein secretion rate and ability to activate myogenesis. The loss of N-glycans in murine GASP-1 leads to a decrease in protein secretion rate but does not affect its ability to activate myogenesis in C2C12 cells.
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