1. Characterisation of class I and II α-mannosidases from Drosophila melanogaster.
- Author
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Nemčovičová I, Šesták S, Rendić D, Plšková M, Mucha J, and Wilson IB
- Subjects
- Amino Acid Sequence, Animals, Drosophila Proteins genetics, Drosophila Proteins metabolism, Kinetics, Molecular Sequence Data, Polysaccharides metabolism, Substrate Specificity, alpha-Mannosidase genetics, alpha-Mannosidase metabolism, Drosophila Proteins chemistry, Drosophila melanogaster enzymology, alpha-Mannosidase chemistry
- Abstract
Homology searches indicated that up to five class I α-mannosidases (glycohydrolase family 47) and eight class II α-mannosidases (glycohydrolase family 38) are encoded by the fruitfly (Drosophila melanogaster) genome. Selected example mannosidases were expressed in secreted form using the yeast Pichia pastoris. A number of characteristics of these enzymes were determined with p-nitrophenyl-α-mannoside as substrate; particularly striking were the low optima (pH 5) of three class II mannosidases most closely related to known lysosomal mannosidases and the distinct Co(II)-requirement of a mannosidase previously named ManIIb. Some of the recombinant mannosidases were demonstrably active towards oligomannosidic glycans, specifically, the Co(II)-requiring ManIIb, two 'acidic' mannosidases and the class I mas-1 mannosidase. Other than previous characterisations of the well-known Golgi mannosidase II, this is the first study summarising various properties of recombinant mannosidases from the fruitfly.
- Published
- 2013
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