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The class I α1,2-mannosidases of Caenorhabditis elegans

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Abstract

During the biosynthesis of N-glycans in multicellular eukaryotes, glycans with the compositions Man5GlcNAc2-3 are key intermediates. However, to reach this ‘decision point’, these N-glycans are first processed from Glc3Man9GlcNAc2 through to Man5GlcNAc2 by a number of glycosidases, whereby up to four α1-2-linked mannose residues are removed by class I mannosidases (glycohydrolase family 47). Whereas in the yeast Saccharomyces cerevisiae there are maximally three members of this protein family, in higher organisms there are multiple class I mannosidases residing in the endoplasmic reticulum and Golgi apparatus. The genome of the model nematode Caenorhabditis elegans encodes seven members of this protein family, whereby four are predicted to be classical processing mannosidases and three are related proteins with roles in quality control. In this study, cDNAs encoding the four predicted mannosidases were cloned and expressed in Pichia pastoris and the activity of these enzymes, designated MANS-1, MANS-2, MANS-3 and MANS-4, was verified. The first two can, dependent on the incubation time, remove three to four residues from Man9GlcNAc2, whereas the action of the other two results in the appearance of the B isomer of Man8GlcNAc2; together the complementary activities of these enzymes result in processing to Man5GlcNAc2. With these data, another gap is closed in our understanding of the N-glycan biosynthesis pathway of the nematode worm.

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Acknowledgements

I thank members of my group, Dr. Peter Both, Natalie Öhl, Alba Hykollari and Dr. Katharina Paschinger, for supporting this work by helping with the expression, tryptic digestion and mass spectrometry experiments.

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  1. Department für Chemie, Universität für Bodenkultur, Muthgasse 18, 1190, Wien, Austria

    Iain B. H. Wilson

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Correspondence to Iain B. H. Wilson.

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Wilson, I.B.H. The class I α1,2-mannosidases of Caenorhabditis elegans . Glycoconj J 29, 173–179 (2012). https://doi.org/10.1007/s10719-012-9378-1

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