Identification of PNGase-dependent ERAD substrates in Saccharomyces cerevisiae

We report herein the identification of endogenous substrates for the cytoplasmic PNGase in Saccharomyces cerevisiae. Using an isotope-coded glycosylation site-specific tagging (IGOT) method-based LC/MS analysis, 11 glycoproteins were specifically detected in the cytosol of PNGase-deletion cells (png1). Among these molecules, at least five glycoproteins were clearly identified as ERAD substrates in vivo. Moreover, four out of the five proteins were found to be either deglycosylated by PNGase in vivo or the overall degradation was delayed in a png1 mutant. Our results clearly indicate that the IGOT method promises to be a powerful tool for the identification of endogenous substrates for the cytoplasmic PNGase.

Source: Biochemical Journal - September 26, 2016 Category: Biochemistry Authors: Hosomi, A., Fujita, M., Tomioka, A., Kaji, H., Suzuki, T. Tags: Research Articles Source Type: research