title: Phosphorylation of arylsulphatase A occurs through multiple interactions with the UDP-N-acetylglucosamine-1-phosphotransferase proximal and distal to its retrieval site by the KDEL receptor.
abstract: Phosphorylation of oligosaccharides of the lysosomal enzyme arylsulphatase A (ASA), which accumulate in the secretions of cells that mis-sort most of the newly synthesized lysosomal enzymes due to a deficiency of mannose 6-phosphate receptors, was found to be site specific. ASA residing within the secretory route of these cells contains about one third of the incorporated [2-3H]mannose in phosphorylated oligosaccharides. Oligosaccharides carrying two phosphate groups are almost 2-fold less frequent than those with one phosphate group and only a few of the phosphate groups are uncovered. Addition of a KDEL (Lys-Asp-Glu-Leu) retention signal prolongs the residence time of ASA within the secretory route 6-fold, but does not result in more efficient phosphorylation. In contrast, more than 90% of the [2-3H]mannose incorporated into secreted ASA (with or without a KDEL retention signal) is present in phosphorylated oligosaccharides. Those with two phosphate groups are almost twice as frequent as those with one phosphate group and most of the phosphate groups are uncovered. Thus, ASA receives N-acetylglucosamine 1-phosphate groups in a sequential manner at two or more sites located within the secretory route proximal and distal to the site where ASA is retrieved by the KDEL receptor, i.e. proximal to the trans-Golgi. At each of these sites up to two N-acetylglucosamine 1-phosphate groups can be added to a single oligosaccharide. Of several drugs known to inhibit transit of ASA through the secretory route only the ionophore monensin had a major inhibitory effect on phosphorylation, uncovering and sialylation.
authors.affiliation: Georg-August-Universität, Abteilung Biochemie II, Gosslerstrasse 12d, 37073 Göttingen, Germany.
authors.forename: F
authors.initials: F
authors.lastname: Dittmer
authors.forename: K
authors.initials: K
authors.lastname: von Figura
mesh_terms: D000117:Acetylglucosamine / Q000031:analogs & derivatives / Q000378:metabolism; D000643:Ammonium Chloride / Q000494:pharmacology; D002553:Cerebroside-Sulfatase / Q000235:genetics / Q000378:metabolism*; D002738:Chloroquine / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D005947:Glucose / Q000378:metabolism; D006031:Glycosylation; D006056:Golgi Apparatus / Q000187:drug effects / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008247:Lysosomes / Q000201:enzymology; D008358:Mannose / Q000378:metabolism; D008360:Mannosephosphates / Q000378:metabolism; D008985:Monensin / Q000494:pharmacology; D019158:N-Acetylneuraminic Acid / Q000378:metabolism; D009844:Oligosaccharides / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D011319:Primaquine / Q000494:pharmacology; D021382:Protein Sorting Signals / Q000235:genetics / Q000378:metabolism; D017527:Receptor, IGF Type 2 / Q000172:deficiency / Q000235:genetics / Q000502:physiology; D018000:Receptors, Peptide / Q000378:metabolism*; D017855:Transferases (Other Substituted Phosphate Groups) / Q000378:metabolism*
chemical_list: C064459:KDEL receptor; D008360:Mannosephosphates; D009844:Oligosaccharides; D021382:Protein Sorting Signals; D017527:Receptor, IGF Type 2; D018000:Receptors, Peptide; D000643:Ammonium Chloride; C027693:mannose-6-phosphate; C026146:N-acetylglucosamine-1-phosphate; D002738:Chloroquine; D008985:Monensin; D017855:Transferases (Other Substituted Phosphate Groups); C030012:UDP-N-acetylglucosamine-lysosomal-enzyme-N-acetylglucosaminephosphotransferase; D002553:Cerebroside-Sulfatase; D019158:N-Acetylneuraminic Acid; D005947:Glucose; D011319:Primaquine; D008358:Mannose; D000117:Acetylglucosamine