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Expression, purification and preliminary crystallographic analysis of Drosophila melanogaster lysosomal ?-mannosidase.

ABSTRACT: The lysosomal ?-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 ?-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.

SUBMITTER: Nemcovicova I

PROVIDER: S-EPMC3412785 | biostudies-literature | 2012 Aug

REPOSITORIES: biostudies-literature

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Expression, purification and preliminary crystallographic analysis of Drosophila melanogaster lysosomal α-mannosidase.

Nemčovičová I I Nemčovič M M Sesták S S Plšková M M Wilson I B H IB Mucha J J

Acta crystallographica. Section F, Structural biology and crystallization communications 20120731 Pt 8

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 wa ...[more]

PMID: 22869134

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