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Evolutionary diversity of social amoebae N-glycomes may support interspecific autonomy

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Abstract

Multiple species of cellular slime mold (CSM) amoebae share overlapping subterranean environments near the soil surface. Despite similar life-styles, individual species form independent starvation-induced fruiting bodies whose spores can renew the life cycle. N-glycans associated with the cell surface glycocalyx have been predicted to contribute to interspecific avoidance, resistance to pathogens, and prey preference. N-glycans from five CSM species that diverged 300–600 million years ago and whose genomes have been sequenced were fractionated into neutral and acidic pools and profiled by MALDI-TOF-MS. Glycan structure models were refined using linkage specific antibodies, exoglycosidase digestions, MALDI-MS/MS, and chromatographic studies. Amoebae of the type species Dictyostelium discoideum express modestly trimmed high mannose N-glycans variably modified with core α3-linked Fuc and peripherally decorated with 0–2 residues each of β-GlcNAc, Fuc, methylphosphate and/or sulfate, as reported previously. Comparative analyses of D. purpureum, D. fasciculatum, Polysphondylium pallidum, and Actyostelium subglobosum revealed that each displays a distinctive spectrum of high-mannose species with quantitative variations in the extent of these modifications, and qualitative differences including retention of Glc, mannose methylation, and absence of a peripheral GlcNAc, fucosylation, or sulfation. Starvation-induced development modifies the pattern in all species but, except for universally observed increased mannose-trimming, the N-glycans do not converge to a common profile. Correlations with glycogene repertoires will enable future reverse genetic studies to eliminate N-glycomic differences to test their functions in interspecific relations and pathogen evasion.

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Abbreviations

2-AB:

2-aminobenzamide

ACN:

Acetonitrile

CSM:

Cellular slime mold

DTT:

Dithiothreitol

H8N4F:

Notation for a glycan containing 8 hexoses, 4 HexNAc residues, and 1 deoxyHex expected to be fucose

TFA:

Trifluoroacetic acid

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Acknowledgments

This project was partially supported by NIH R01-GM037539 and the OCMG, which received funding from the OUHSC Dept. of Biochemistry & Molecular Biology and the OUHSC VP Office for Research. We are grateful to Jennifer Johnson for her technical assistance, the Dictyostelium Stock Center (Northwestern University) and Adam Kuspa (Baylor) for providing cells, and to Christine Scaman (Univ. of British Columbia) for providing yeast α-glucosidase-I.

Conflict of interest

None declared.

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Author notes

  1. Christa L. Feasley

    Present address: Thermo-Fisher Scientific, West Palm Beach, FL, 33407, USA

Authors and Affiliations

  1. Department of Biochemistry & Molecular Biology, Oklahoma Center for Medical Glycobiology, University of Oklahoma Health Sciences Center, 975 NE 10th St., BRC-415, OUHSC, Oklahoma City, OK, 73104, USA

    Christa L. Feasley, Hanke van der Wel & Christopher M. West

  2. Department of Biochemistry and MolecularBiology, Fred C. Davison Life Sciences Complex, The University of Georgia, Athens, GA, 30602, USA

    Christopher M. West

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Feasley, C.L., van der Wel, H. & West, C.M. Evolutionary diversity of social amoebae N-glycomes may support interspecific autonomy. Glycoconj J 32, 345–359 (2015). https://doi.org/10.1007/s10719-015-9592-8

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