title: Multiple Domains of GlcNAc-1-phosphotransferase Mediate Recognition of Lysosomal Enzymes.
abstract: The Golgi enzyme UDP-GlcNAc:lysosomal enzymeN-acetylglucosamine-1-phosphotransferase (GlcNAc-1-phosphotransferase), an α2β2γ2hexamer, mediates the initial step in the addition of the mannose 6-phosphate targeting signal on newly synthesized lysosomal enzymes. This tag serves to direct the lysosomal enzymes to lysosomes. A key property of GlcNAc-1-phosphotransferase is its unique ability to distinguish the 60 or so lysosomal enzymes from the numerous non-lysosomal glycoproteins with identical Asn-linked glycans. In this study, we demonstrate that the two Notch repeat modules and the DNA methyltransferase-associated protein interaction domain of the α subunit are key components of this recognition process. Importantly, different combinations of these domains are involved in binding to individual lysosomal enzymes. This study also identifies the γ-binding site on the α subunit and demonstrates that in the majority of instances the mannose 6-phosphate receptor homology domain of the γ subunit is required for optimal phosphorylation. These findings serve to explain how GlcNAc-1-phosphotransferase recognizes a large number of proteins that lack a common structural motif.
authors.affiliation: From the Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
authors.forename: Eline
authors.initials: E
authors.lastname: van Meel
authors.affiliation: From the Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
authors.forename: Wang-Sik
authors.initials: WS
authors.lastname: Lee
authors.affiliation: From the Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
authors.forename: Lin
authors.initials: L
authors.lastname: Liu
authors.affiliation: From the Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
authors.forename: Yi
authors.initials: Y
authors.lastname: Qian
authors.affiliation: From the Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
authors.forename: Balraj
authors.initials: B
authors.lastname: Doray
authors.affiliation: From the Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri 63110 skornfel@dom.wustl.edu.
authors.forename: Stuart
authors.initials: S
authors.lastname: Kornfeld
mesh_terms: D000595:Amino Acid Sequence; D017353:Gene Deletion; D006367:HeLa Cells; D006801:Humans; D008247:Lysosomes / Q000201:enzymology* / Q000378:metabolism; D008360:Mannosephosphates / Q000378:metabolism*; D008969:Molecular Sequence Data; D010766:Phosphorylation; D054730:Protein Interaction Domains and Motifs; D051880:Receptors, Notch / Q000737:chemistry / Q000378:metabolism; D017855:Transferases (Other Substituted Phosphate Groups) / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D008360:Mannosephosphates; D051880:Receptors, Notch; C027693:mannose-6-phosphate; D017855:Transferases (Other Substituted Phosphate Groups); C505859:alpha-beta GlcNAc-1-phosphotransferase, human
keywords: lysosomal glycoprotein; lysosomal storage disease; lysosome; mannose 6-phosphate (Man-6-P); phosphorylation
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