title: Golgi apparatus immunolocalization of endomannosidase suggests post-endoplasmic reticulum glucose trimming: implications for quality control.
abstract: Trimming of N-linked oligosaccharides by endoplasmic reticulum (ER) glucosidase II is implicated in quality control of protein folding. An alternate glucosidase II-independent deglucosylation pathway exists, in which endo-alpha-mannosidase cleaves internally the glucose-substituted mannose residue of oligosaccharides. By immunogold labeling, we detected most endomannosidase in cis/medial Golgi cisternae (83.8% of immunogold labeling) and less in the intermediate compartment (15.1%), but none in the trans-Golgi apparatus and ER, including its transitional elements. This dual localization became more pronounced under 15 degrees C conditions indicative of two endomannosidase locations. Under experimental conditions when the intermediate compartment marker p58 was retained in peripheral sites, endomannosidase was redistributed to the Golgi apparatus. Double immunogold labeling established a mutually exclusive distribution of endomannosidase and glucosidase II, whereas calreticulin was observed in endomannosidase-reactive sites (17.3% in intermediate compartment, 5.7% in Golgi apparatus) in addition to the ER (77%). Our results demonstrate that glucose trimming of N-linked oligosaccharides is not limited to the ER and that protein deglucosylation by endomannosidase in the Golgi apparatus and intermediate compartment additionally ensures that processing to mature oligosaccharides can continue. Thus, endomannosidase localization suggests that a quality control of N-glycosylation exists in the Golgi apparatus.
authors: Division of Cell and Molecular Pathology, Department of Pathology, University of Zürich, CH-8091 Zürich, Switzerland.
authors: C
authors: C
authors: Zuber
authors: M J
authors: MJ
authors: Spiro
authors: B
authors: B
authors: Guhl
authors: R G
authors: RG
authors: Spiro
authors: J
authors: J
authors: Roth
mesh_terms: D000818:Animals; D002135:Calcium-Binding Proteins / Q000378:metabolism; D037282:Calreticulin; D002451:Cell Compartmentation; D002460:Cell Line; D004721:Endoplasmic Reticulum / Q000201:enzymology* / Q000648:ultrastructure; D005947:Glucose / Q000378:metabolism*; D006031:Glycosylation; D006056:Golgi Apparatus / Q000201:enzymology* / Q000648:ultrastructure; D008099:Liver / Q000201:enzymology; D037241:Mannose-Binding Lectins; D008361:Mannosidases / Q000276:immunology / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D011499:Protein Processing, Post-Translational; D051381:Rats; D012261:Ribonucleoproteins / Q000378:metabolism; D000520:alpha-Glucosidases / Q000378:metabolism
chemical_list: D002135:Calcium-Binding Proteins; D037282:Calreticulin; C501458:Lman1 protein, rat; D037241:Mannose-Binding Lectins; D008565:Membrane Proteins; D012261:Ribonucleoproteins; C034251:4-nitrophenyl-alpha-glucosidase; D008361:Mannosidases; C051635:endo-alpha-D-mannosidase; D000520:alpha-Glucosidases; D005947:Glucose