title: Synthetic phospho-oligosaccharide fragments of lipophosphoglycan as acceptors for Leishmania major alpha-D-mannosylphosphate transferase.
abstract: Protozoan parasites of the genus Leishmania synthesise lipophosphoglycans, phosphoglycans and proteophosphoglycans that contain phosphosaccharide-repeat units of [-6Gal beta 1-4Man alpha 1-P-]. In this study, a GDP-Man-dependent alpha-mannosylphosphate-transferase activity was detected in washed Leishmania major membranes using synthetic phospho-oligosaccharide fragments of lipophosphoglycan as acceptor substrates. The divalent-cation-dependent alpha-mannosylphosphate-transferase activity had an apparent K(m) for GDP-Man of about 15-20 microM and a pH optimum of 7.0. The activity showed a requirement for a non-reducing terminal beta Gal residue and for one or more phosphodiester units preceding the acceptor site. Based on these results, the activity may be defined as a GDP-Man: Gal beta 1-4Man alpha 1-P-R alpha-mannosylphosphate-transferase. This acceptor specificity is consistent with a role for the alpha-mannosylphosphate transferase in the elongation of phosphosaccharide-repeat domains of Leishmania glycoconjugates rather than in the priming of these domains. An identical or similar activity must exist in the amastigote forms of the Leishmania that produce and secrete proteophosphoglycan material and the activity therefore represents a feasible target for the development of chemotherapeutics.
authors.affiliation: Department of Biochemistry, University of Dundee, Scotland.
authors.forename: G M
authors.initials: GM
authors.lastname: Brown
authors.forename: A R
authors.initials: AR
authors.lastname: Millar
authors.forename: C
authors.initials: C
authors.lastname: Masterson
authors.forename: J S
authors.initials: JS
authors.lastname: Brimacombe
authors.forename: A V
authors.initials: AV
authors.lastname: Nikolaev
authors.forename: M A
authors.initials: MA
authors.lastname: Ferguson
mesh_terms: D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002417:Cattle; D006028:Glycosphingolipids / Q000737:chemistry* / Q000378:metabolism*; D007295:Inositol Phosphates; D007700:Kinetics; D018320:Leishmania major / Q000378:metabolism*; D008297:Male; D008364:Mannosyltransferases / Q000378:metabolism*; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*; D011134:Polysaccharides; D013379:Substrate Specificity; D013737:Testis; D001616:beta-Galactosidase
chemical_list: D006028:Glycosphingolipids; D007295:Inositol Phosphates; D009844:Oligosaccharides; D011134:Polysaccharides; C028039:inositol phosphate glycan; C008290:lipophosphonoglycan; D008364:Mannosyltransferases; C022755:dolichyl-phosphate beta-D-mannosyltransferase; D001616:beta-Galactosidase