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

• Published in: Acta crystallographica. Section F, Structural biology and crystallization communications Vol. 68; no. 8; pp. 965 - 970
• Main Authors: Nemčovičová, I., Nemčovič, M., Šesták, S., Plšková, M., Wilson, I. B. H., Mucha, J.
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England International Union of Crystallography 01.08.2012
• Subjects: alpha-Mannosidase - chemistry; alpha-Mannosidase - genetics; alpha-Mannosidase - isolation & purification; Animals; Crystallization Communications; Crystallography; Crystallography, X-Ray; Crystals; Degradation; Diffraction; Drosophila; Drosophila melanogaster; Drosophila melanogaster - enzymology; Gene Expression; glycosyl hydrolase family 38; lysosomal α-mannosidases; Peptides; Recombinant; Similarity
• ISSN: 1744-3091; 1744-3091
• DOI: 10.1107/S1744309112029375

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 P212121 and diffracted to a minimum d‐spacing of 3.5 Å.