Multiple O-glycoforms on the spore coat protein sp96 in Dictyostelium discoideum. Fuc(α1-3)GlcNAc-α-1-P-Ser is the major modification

Marcus Mreyen, Alan Champion, Supriya Srinivasan, Peter Karuso, Keith L. Williams, Nicolle H. Packer

Research output: Contribution to journalArticleResearchpeer-review

Abstract

A decreased level of fucosylation on certain spore coat proteins of Dictyostelium discoideum alters the permeability of the spore coat. Here the post-translational modifications of a major spore coat protein, SP96, are studied in a wild type strain (X22) and a fucosylation-defective mutant (HU2470). A novel phosphoglycan structure on SP96 of the wild type strain, consisting of Fuc(α1-3)GlcNAc-α-1-P-Ser was identified by electrospray ionization mass spectrometry and NMR. It was shown using monosaccharide and gas chromatography mass spectrometry analysis that SP96 in the mutant HU2470 contained approximately 20% of wild type levels of fucose, as a result of a missing terminal fucose on the novel glycan structure. The results support previous predictions, based on inhibition studies on different fucose- deficient strains, about the nature of monoclonal antibody epitopes identified by monoclonal antibodies MUD62 and MUD166, which are known to identify O-linked glycans (Champion, A., Griffiths, K., Gooley, A. A., Gonzalez, B. Y., Gritzali, M., West, C. M., and Williams, K. L. (1995) Microbiology 141, 785-797). Quantitative studies on wild type SP96 indicated that there were approximately 60 sites with phosphodiester-linked N- acetylglucosamine-fucose disaccharide units and a further approximately 20 sites with fucose directly linked to the protein. Over 70% of the serine sites are modified, with less than 1% of these sites as phosphoserine. Threonine and tyrosine residues were not found to be modified.

LanguageEnglish
Pages12164-12174
Number of pages11
JournalJournal of Biological Chemistry
Volume275
Issue number16
DOIs
Publication statusPublished - 21 Apr 2000

Fingerprint

Dictyostelium
Fucose
Capsid Proteins
Spores
Mass spectrometry
Polysaccharides
Monoclonal Antibodies
Phosphoserine
Microbiology
Electrospray ionization
Acetylglucosamine
Electrospray Ionization Mass Spectrometry
Monosaccharides
Disaccharides
Threonine
Post Translational Protein Processing
Gas chromatography
Gas Chromatography-Mass Spectrometry
Serine
Tyrosine

Cite this

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title = "Multiple O-glycoforms on the spore coat protein sp96 in Dictyostelium discoideum. Fuc(α1-3)GlcNAc-α-1-P-Ser is the major modification",
abstract = "A decreased level of fucosylation on certain spore coat proteins of Dictyostelium discoideum alters the permeability of the spore coat. Here the post-translational modifications of a major spore coat protein, SP96, are studied in a wild type strain (X22) and a fucosylation-defective mutant (HU2470). A novel phosphoglycan structure on SP96 of the wild type strain, consisting of Fuc(α1-3)GlcNAc-α-1-P-Ser was identified by electrospray ionization mass spectrometry and NMR. It was shown using monosaccharide and gas chromatography mass spectrometry analysis that SP96 in the mutant HU2470 contained approximately 20{\%} of wild type levels of fucose, as a result of a missing terminal fucose on the novel glycan structure. The results support previous predictions, based on inhibition studies on different fucose- deficient strains, about the nature of monoclonal antibody epitopes identified by monoclonal antibodies MUD62 and MUD166, which are known to identify O-linked glycans (Champion, A., Griffiths, K., Gooley, A. A., Gonzalez, B. Y., Gritzali, M., West, C. M., and Williams, K. L. (1995) Microbiology 141, 785-797). Quantitative studies on wild type SP96 indicated that there were approximately 60 sites with phosphodiester-linked N- acetylglucosamine-fucose disaccharide units and a further approximately 20 sites with fucose directly linked to the protein. Over 70{\%} of the serine sites are modified, with less than 1{\%} of these sites as phosphoserine. Threonine and tyrosine residues were not found to be modified.",
author = "Marcus Mreyen and Alan Champion and Supriya Srinivasan and Peter Karuso and Williams, {Keith L.} and Packer, {Nicolle H.}",
year = "2000",
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Multiple O-glycoforms on the spore coat protein sp96 in Dictyostelium discoideum. Fuc(α1-3)GlcNAc-α-1-P-Ser is the major modification. / Mreyen, Marcus; Champion, Alan; Srinivasan, Supriya; Karuso, Peter; Williams, Keith L.; Packer, Nicolle H.

In: Journal of Biological Chemistry, Vol. 275, No. 16, 21.04.2000, p. 12164-12174.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Mreyen, Marcus

AU - Champion, Alan

AU - Srinivasan, Supriya

AU - Karuso, Peter

AU - Williams, Keith L.

AU - Packer, Nicolle H.

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N2 - A decreased level of fucosylation on certain spore coat proteins of Dictyostelium discoideum alters the permeability of the spore coat. Here the post-translational modifications of a major spore coat protein, SP96, are studied in a wild type strain (X22) and a fucosylation-defective mutant (HU2470). A novel phosphoglycan structure on SP96 of the wild type strain, consisting of Fuc(α1-3)GlcNAc-α-1-P-Ser was identified by electrospray ionization mass spectrometry and NMR. It was shown using monosaccharide and gas chromatography mass spectrometry analysis that SP96 in the mutant HU2470 contained approximately 20% of wild type levels of fucose, as a result of a missing terminal fucose on the novel glycan structure. The results support previous predictions, based on inhibition studies on different fucose- deficient strains, about the nature of monoclonal antibody epitopes identified by monoclonal antibodies MUD62 and MUD166, which are known to identify O-linked glycans (Champion, A., Griffiths, K., Gooley, A. A., Gonzalez, B. Y., Gritzali, M., West, C. M., and Williams, K. L. (1995) Microbiology 141, 785-797). Quantitative studies on wild type SP96 indicated that there were approximately 60 sites with phosphodiester-linked N- acetylglucosamine-fucose disaccharide units and a further approximately 20 sites with fucose directly linked to the protein. Over 70% of the serine sites are modified, with less than 1% of these sites as phosphoserine. Threonine and tyrosine residues were not found to be modified.

AB - A decreased level of fucosylation on certain spore coat proteins of Dictyostelium discoideum alters the permeability of the spore coat. Here the post-translational modifications of a major spore coat protein, SP96, are studied in a wild type strain (X22) and a fucosylation-defective mutant (HU2470). A novel phosphoglycan structure on SP96 of the wild type strain, consisting of Fuc(α1-3)GlcNAc-α-1-P-Ser was identified by electrospray ionization mass spectrometry and NMR. It was shown using monosaccharide and gas chromatography mass spectrometry analysis that SP96 in the mutant HU2470 contained approximately 20% of wild type levels of fucose, as a result of a missing terminal fucose on the novel glycan structure. The results support previous predictions, based on inhibition studies on different fucose- deficient strains, about the nature of monoclonal antibody epitopes identified by monoclonal antibodies MUD62 and MUD166, which are known to identify O-linked glycans (Champion, A., Griffiths, K., Gooley, A. A., Gonzalez, B. Y., Gritzali, M., West, C. M., and Williams, K. L. (1995) Microbiology 141, 785-797). Quantitative studies on wild type SP96 indicated that there were approximately 60 sites with phosphodiester-linked N- acetylglucosamine-fucose disaccharide units and a further approximately 20 sites with fucose directly linked to the protein. Over 70% of the serine sites are modified, with less than 1% of these sites as phosphoserine. Threonine and tyrosine residues were not found to be modified.

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