Endomannosidase processes oligosaccharides of alpha1-antitrypsin and its naturally occurring genetic variants in the Golgi apparatus.
Endomannosidase provides an alternate glucose-trimming pathway in the Golgi apparatus. However, it is unknown if the action of endomannosidase is dependent on the conformation of the substrate. We have investigated the processing by endomannosidase of the alpha1-antitrypsin oligosaccharides and its disease-causing misfolded Z and Hong Kong variants. Oligosaccharides of wild-type and misfolded alpha1-antitrypsin expressed in castanospermine-treated hepatocytes or glucosidase II-deficient Phar 2.7 cells were selectively processed by endomannosidase and subsequently converted to complex type oligosaccharides as indicated by Endo H resistance and PNGase F sensitivity. Overexpression of endomannosidase in castanospermine-treated hepatocytes resulted in processing of all oligosaccharides of wild-type and variants of alpha1-antitrypsin. Thus, endomannosidase does not discriminate the folding state of the substrate and provides a back-up mechanism for completion of N-glycosylation of endoplasmic reticulum-escaped glucosylated glycoproteins. For exported misfolded glycoproteins, this would provide a pathway for the formation of mature oligosaccharides important for their proper trafficking and correct functioning.
Torossi
T
T
Department of Pathology, Division of Cell and Molecular Pathology, University of Zurich, Schmelzbergstrasse 12, 8091, Zurich, Switzerland.
Fan
J-Y
JY
Sauter-Etter
K
K
Roth
J
J
Ziak
M
M
D000818:Animals; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D017931:DNA Primers; D014644:Genetic Variation; D006026:Glycoside Hydrolases; D006031:Glycosylation; D006056:Golgi Apparatus / Q000235:genetics* / Q000378:metabolism*; D008223:Lymphoma; D008361:Mannosidases / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D016297:Mutagenesis, Site-Directed; D009844:Oligosaccharides / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D000515:alpha 1-Antitrypsin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D017931:DNA Primers; D008565:Membrane Proteins; D009844:Oligosaccharides; D011994:Recombinant Proteins; D000515:alpha 1-Antitrypsin; D006026:Glycoside Hydrolases; D008361:Mannosidases; C505101:alpha-1-2-endomannosidase, rat