title: Expression, purification and preliminary crystallographic analysis of Drosophila melanogaster lysosomal α-mannosidase.
abstract: The lysosomal α-mannosidases are class II mannosidases that belong to glycoside hydrolase family 38 and play an important role in the degradation of asparagine-linked carbohydrates of glycoproteins. Based on peptide similarity to human and bovine lysosomal mannosidase (LM), recombinant α-mannosidase from Drosophila melanogaster (dLM408) was cloned and heterologously expressed in Pichia pastoris. The recombinant form of dLM408 designed for structural analysis lacks the transmembrane domain and was crystallized using standard vapour-diffusion and counter-diffusion techniques. The crystals grew as flat plates and as tetragonal bipyramids, respectively. The plate-shaped crystals exhibited the symmetry of space group P2(1)2(1)2(1) and diffracted to a minimum d-spacing of 3.5 Å.
authors.affiliation: Department of Cellular Biology, La Jolla Institute for Allergy and Immunology, 9420 Athena Circle, La Jolla, CA 92037, USA.
authors.forename: I
authors.initials: I
authors.lastname: Nemčovičová
authors.forename: M
authors.initials: M
authors.lastname: Nemčovič
authors.forename: S
authors.initials: S
authors.lastname: Sesták
authors.forename: M
authors.initials: M
authors.lastname: Plšková
authors.forename: I B H
authors.initials: IB
authors.lastname: Wilson
authors.forename: J
authors.initials: J
authors.lastname: Mucha
mesh_terms: D000818:Animals; D018360:Crystallography, X-Ray; D004331:Drosophila melanogaster / Q000201:enzymology*; D015870:Gene Expression; D043323:alpha-Mannosidase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification
chemical_list: D043323:alpha-Mannosidase