title: Temporal association of the N- and O-linked glycosylation events and their implication in the polarized sorting of intestinal brush border sucrase-isomaltase, aminopeptidase N, and dipeptidyl peptidase IV.
abstract: The temporal association between O-glycosylation and processing of N-linked glycans in the Golgi apparatus as well as the implication of these events in the polarized sorting of three brush border proteins has been the subject of the current investigation. O-Glycosylation of pro-sucrase-isomaltase (pro-SI), aminopeptidase N (ApN), and dipeptidyl peptidase IV (DPPIV) is drastically reduced when processing of the mannose-rich N-linked glycans is blocked by deoxymannojirimycin, an inhibitor of the Golgi-located mannosidase I. By contrast, O-glycosylation is not affected in the presence of swainsonine, an inhibitor of Golgi mannosidase II. The results indicate that removal of the outermost mannose residues by mannosidase I from the mannose-rich N-linked glycans is required before O-glycosylation can ensue. On the other hand, subsequent mannose residues in the core chain impose no sterical constraints on the progression of O-glycosylation. Reduction or modification of N- and O-glycosylation do not affect the transport of pro-SI, ApN, or DPPIV to the cell surface per se. However, the polarized sorting of two of these proteins, pro-SI and DPPIV, to the apical membrane is substantially altered when O-glycans are not completely processed, while the sorting of ApN is not affected. The processing of N-linked glycans, on the other hand, has no influence on sorting of all three proteins. The results indicate that O-linked carbohydrates are at least a part of the sorting mechanism of pro-SI and DPPIV. The sorting of ApN implicates neither O-linked nor N-linked glycans and is driven most likely by carbohydrate-independent mechanisms.
authors.affiliation: Department of Physiological Chemistry, School of Veterinary Medicine Hannover, Bünteweg 17, D-30559 Hannover, Germany. hnaim@biochemie.tiho-hannover.de
authors.forename: H Y
authors.initials: HY
authors.lastname: Naim
authors.forename: G
authors.initials: G
authors.lastname: Joberty
authors.forename: M
authors.initials: M
authors.lastname: Alfalah
authors.forename: R
authors.initials: R
authors.lastname: Jacob
mesh_terms: D017485:1-Deoxynojirimycin / Q000494:pharmacology; D018826:CD13 Antigens / Q000378:metabolism*; D018938:Caco-2 Cells; D018819:Dipeptidyl Peptidase 4 / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006031:Glycosylation / Q000187:drug effects; D006596:Hexosaminidases / Q000378:metabolism; D006801:Humans; D007422:Intestines / Q000201:enzymology*; D008361:Mannosidases / Q000037:antagonists & inhibitors; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / Q000378:metabolism; D008871:Microvilli / Q000201:enzymology*; D011134:Polysaccharides / Q000378:metabolism; D013394:Sucrase-Isomaltase Complex / Q000378:metabolism*; D017026:Swainsonine / Q000494:pharmacology
chemical_list: D004791:Enzyme Inhibitors; D011134:Polysaccharides; D017485:1-Deoxynojirimycin; D006596:Hexosaminidases; D008361:Mannosidases; D013394:Sucrase-Isomaltase Complex; C046090:mannosyl-oligosaccharide 1,2-alpha-mannosidase; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D018826:CD13 Antigens; D018819:Dipeptidyl Peptidase 4; D017026:Swainsonine