title: Man2C1, an alpha-mannosidase, is involved in the trimming of free oligosaccharides in the cytosol.
abstract: The endoplasmic-reticulum-associated degradation of misfolded (glyco)proteins ensures that only functional, correctly folded proteins exit from the endoplasmic reticulum and that misfolded ones are degraded by the ubiquitin-proteasome system. During the degradation of misfolded glycoproteins, they are deglycosylated by the PNGase (peptide:N-glycanase). The free oligosaccharides released by PNGase are known to be further catabolized by a cytosolic alpha-mannosidase, although the gene encoding this enzyme has not been identified unequivocally. The findings in the present study demonstrate that an alpha-mannosidase, Man2C1, is involved in the processing of free oligosaccharides that are formed in the cytosol. When the human Man2C1 orthologue was expressed in HEK-293 cells, most of the enzyme was localized in the cytosol. Its activity was enhanced by Co2+, typical of other known cytosolic alpha-mannosidases so far characterized from animal cells. The down-regulation of Man2C1 activity by a small interfering RNA drastically changed the amount and structure of oligosaccharides accumulating in the cytosol, demonstrating that Man2C1 indeed is involved in free oligosaccharide processing in the cytosol. The oligosaccharide processing in the cytosol by PNGase, endo-beta-N-acetylglucosaminidase and alpha-mannosidase may represent the common 'non-lysosomal' catabolic pathway for N-glycans in animal cells, although the molecular mechanism as well as the functional importance of such processes remains to be determined.
authors.affiliation: Department of Biochemistry, Osaka University Graduate School of Medicine, Osaka, 565-0871, Japan. tsuzuki@biochem.med.osaka-u.ac.jp
authors.forename: Tadashi
authors.initials: T
authors.lastname: Suzuki
authors.forename: Izumi
authors.initials: I
authors.lastname: Hara
authors.forename: Miyako
authors.initials: M
authors.lastname: Nakano
authors.forename: Masaki
authors.initials: M
authors.lastname: Shigeta
authors.forename: Takatoshi
authors.initials: T
authors.lastname: Nakagawa
authors.forename: Akihiro
authors.initials: A
authors.lastname: Kondo
authors.forename: Yoko
authors.initials: Y
authors.lastname: Funakoshi
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Taniguchi
mesh_terms: D000470:Alkaloids / Q000494:pharmacology; D015153:Blotting, Western; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D003035:Cobalt / Q000494:pharmacology; D003600:Cytosol / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D008361:Mannosidases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism; D009842:Oligopeptides; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism*; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D017026:Swainsonine / Q000494:pharmacology; D014162:Transfection; D043323:alpha-Mannosidase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
chemical_list: D000470:Alkaloids; D008562:Membrane Glycoproteins; D009842:Oligopeptides; D009844:Oligosaccharides; D010455:Peptides; D011993:Recombinant Fusion Proteins; C065629:kifunensine; D049452:Green Fluorescent Proteins; D003035:Cobalt; C085706:FLAG peptide; D008361:Mannosidases; C511245:MAN2C1 protein, human; D043323:alpha-Mannosidase; C018021:cobaltous chloride; D017026:Swainsonine