Secretome analysis to elucidate metalloprotease ‐dependent ectodomain shedding of glycoproteins during neuronal differentiation

In this study, to elucidate the ectodomain shedding events that occur during neuronal differentiation, we establish a strategy for quantitative secretomics of glycoproteins released from differentiating neuroblastoma cells into culture medium with or without GM6001, a broad‐spectrum metalloprotease inhibitor. Considering that most of transmembrane and secreted proteins are N‐glycosylated, we include a process of N‐glycosylated peptides enrichment as well as isotope tagging in our secretomics workflow. Our results show that differentiating N1E‐115 neurons secrete numerous glycosylated polypeptides in metalloprotease‐dependent manners. They are derived from cell adhesion molecules such as NCAM1, CADM1, L1CAM, various transporters and receptor proteins. These results show the landscape of ectodomain shedding and other secretory events in differentiating neurons and/or during axon elongation, which should help elucidate the mechanism of neurogenesis and the pathogenesis of neurological disorders. We succeeded in the establishment of a sensitive and reliable method to systematically screen glycoproteins that are secreted from differentiating neurons through metalloprotease‐dependent ectodomain shedding. Our results show the landscape of ectodomain shedding in differentiating neurons, which should help elucidate the mechanisms of neurogenesis and the pathogeneses of neurological disorders.
Source: Genes to Cells - Category: Genetics & Stem Cells Authors: Tags: Brief Report Source Type: research