title: Phosphorylation of the human 46-kDa mannose 6-phosphate receptor in the cytoplasmic domain at serine 56.
abstract: The human 46-kDa mannose 6-phosphate receptor (MPR46) is phosphorylated in its cytoplasmic domain at serine residues. Substitution of cytoplasmic serines (at positions 35 and 56) with alanine, expression of mutant receptors in baby hamster kidney cells, and phosphopeptide mapping revealed that serine 56 is phosphorylated. Mutant MPR46 and wild-type MPR46 were found to be similarly distributed between the cell surface and intracellular membranes. Phosphate incorporation in the presence of cycloheximide indicates that phosphorylation occurred on pre-existing MPR46. Similar half-lives for the wild-type and mutant receptor proteins (approximately 43 h) and the receptor-associated phosphate (1.4 h) were found. The mutant receptors were internalized at the same rate as the wild-type receptors. Expression of mutant MPR46 and wild-type MPR46 in mouse L-cells deficient in 300-kDa mannose 6-phosphate receptors did not affect the sorting of newly synthesized cathepsin D to lysosomes. Phosphorylation of cytoplasmic serine 56 is therefore essential neither for stability nor for cell-surface expression and transport activities of MPR46.
authors.affiliation: Georg-August-Universität Göttingen, Institute of Biochemistry II, Germany.
authors.forename: F
authors.initials: F
authors.lastname: Hemer
authors.forename: C
authors.initials: C
authors.lastname: Körner
authors.forename: T
authors.initials: T
authors.lastname: Braulke
mesh_terms: D000818:Animals; D002402:Cathepsin D / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D003513:Cycloheximide / Q000494:pharmacology; D003593:Cytoplasm / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D007739:L Cells; D051379:Mice; D016297:Mutagenesis, Site-Directed; D010766:Phosphorylation; D011233:Precipitin Tests; D017527:Receptor, IGF Type 2 / Q000737:chemistry / Q000378:metabolism*; D012694:Serine / Q000378:metabolism*; D014357:Trypsin
chemical_list: D017527:Receptor, IGF Type 2; D012694:Serine; D003513:Cycloheximide; D014357:Trypsin; D002402:Cathepsin D