Show legend Show as text

Structure type: oligomer ; 2296 [M+Na]+
Aglycon: s-gp130 peptide
Compound class: N-glycan
Contained glycoepitopes: IEDB_123886,IEDB_123888,IEDB_130701,IEDB_135813,IEDB_136045,IEDB_136104,IEDB_137340,IEDB_137485,IEDB_140116,IEDB_141793,IEDB_141807,IEDB_141828,IEDB_141829,IEDB_141830,IEDB_141831,IEDB_142489,IEDB_143632,IEDB_144562,IEDB_144983,IEDB_145669,IEDB_148493,IEDB_150092,IEDB_151079,IEDB_151531,IEDB_152206,IEDB_152214,IEDB_153212,IEDB_153220,IEDB_164174,IEDB_174333,IEDB_187201,IEDB_429156,IEDB_540671,IEDB_548907,IEDB_857734,IEDB_983930,SB_136,SB_191,SB_196,SB_197,SB_198,SB_33,SB_44,SB_53,SB_67,SB_72,SB_73,SB_74,SB_77,SB_85,SB_86

• Article ID: 5922
  Feasley C, Johnson JM, West CM, Chia CP "Glycopeptidome of a Heavily N-Glycosylated Cell Surface Glycoprotein of Dictyostelium Implicated in Cell Adhesion" - Journal of Proteome Research 9(7) (2010) 3495-3510
  

  Genetic analysis has implicated the cell surface glycoprotein gp130 in cell interactions of the social amoeba Dictyostelium, and information about the utilization of the 18 N-glycosylation sequons present in gp130 is needed to identify critical molecular determinants of its activity. Various glycomics strategies, including mass spectrometry of native and derivatized glycans, monosaccharide analysis, exoglycosidase digestion, and antibody binding, were applied to characterize a nonanchored version secreted from Dictyostelium. s-gp130 is modified by a predominant Man(8)GlcNAc(4) species containing bisecting and intersecting GlcNAc residues and additional high-mannose N-glycans substituted with sulfate, methyl-phosphate, and/or core alpha 3-fucose. Site mapping confirmed the occupancy of 15 sequons, some variably, and glycopeptide analysis confirmed 14 sites and revealed extensive heterogeneity at most sites. Glycopeptide glycoforms ranged from Man(6) to Man(9), GlcNAc(0-2) (peripheral), Fuc(0-2) (including core alpha 3 and peripheral), (SO(4))(0-1), and (MePO(4))(0-1), which represented elements of virtually the entire known cellular N-glycome as inferred from prior metabolic labeling and mass spectrometry studies. gp130, and a family of 14 related predicted glycoproteins whose polypeptide sequences are rapidly diverging in the Dictyostelium lineage, may contribute a functionally important shroud of high-mannose N-glycans at the interface of the amoebae with each other, their predators and prey, and the soil environment.

  mass spectrometry, glycobiology, N-glycan, Dictyostelium, glycopeptidome, gp130

  NCBI PubMed ID: 20443635
  Publication DOI: 10.1021/pr901195c
  Journal NLM ID: 101128775
  Publisher: Washington, DC: American Chemical Society
  Correspondence: Cwest2@ouhsc.edu
  Institutions: Department of Biochemistry & Molecular Biology and Oklahoma Center for Medical Glycobiology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA
  Methods: gel filtration, SDS-PAGE, sugar analysis, Western blotting, amino acid analysis, MALDI-TOF MS, genetic methods, enzymatic digestion, HPAEC-PAD, permethylation, cloning, RP-HPLC, MALDI-TOF/TOF MS, reductive amination, nanoLC-MS/MS
  
   
  
  Dictyostelium discoideum HL250, Dictyostelium discoideum HW11
  CSDB ID 4899 (all data & tools)

Show legend Show as text

Structure type: oligomer ; 2295.8 [M+Na]+
Compound class: N-glycan
Contained glycoepitopes: IEDB_123886,IEDB_123888,IEDB_130701,IEDB_135813,IEDB_136045,IEDB_136104,IEDB_137340,IEDB_137485,IEDB_140116,IEDB_141793,IEDB_141807,IEDB_141828,IEDB_141829,IEDB_141831,IEDB_142489,IEDB_143632,IEDB_144562,IEDB_144983,IEDB_145669,IEDB_148493,IEDB_150092,IEDB_151079,IEDB_151531,IEDB_152206,IEDB_152214,IEDB_153212,IEDB_153220,IEDB_164174,IEDB_174333,IEDB_187201,IEDB_429156,IEDB_548907,IEDB_857734,IEDB_983930,SB_136,SB_191,SB_196,SB_197,SB_198,SB_33,SB_44,SB_53,SB_67,SB_72,SB_73,SB_74,SB_77,SB_85,SB_86

• Article ID: 5923
  Feasley C, van der Wel H, West CM "Evolutionary diversity of social amoebae N-glycomes may support interspecific autonomy" - Glycoconjugate Journal 32(6) (2015) 345-359
  

  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.

  evolution, N-glycan, Dictyostelium, Cellular slime molds, Social amoebae

  NCBI PubMed ID: 25987342
  Publication DOI: 10.1007/s10719-015-9592-8
  Journal NLM ID: 8603310
  Publisher: Kluwer Academic Publishers
  Correspondence: Christopher M. West ; Christopher M. West
  Institutions: 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
  Methods: SDS-PAGE, Western blotting, MALDI-TOF MS, enzymatic digestion, permethylation, column chromatography, MALDI-TOF/TOF MS, bioinformatic analysis (BLASTp)
  
   
  
  Dictyostelium discoideum AX3
  CSDB ID 4492 (all data & tools)

Show legend Show as text

Structure type: oligomer ; 1459 [M+H]+
Aglycon: 2-aminopyridine (PA)
Compound class: N-glycan
Contained glycoepitopes: IEDB_123886,IEDB_123888,IEDB_130701,IEDB_135813,IEDB_136045,IEDB_136104,IEDB_137340,IEDB_137485,IEDB_140116,IEDB_141793,IEDB_141807,IEDB_142489,IEDB_143632,IEDB_144562,IEDB_144983,IEDB_145669,IEDB_148493,IEDB_150092,IEDB_151531,IEDB_152206,IEDB_152214,IEDB_153212,IEDB_164174,IEDB_174333,IEDB_548907,IEDB_983930,SB_136,SB_196,SB_197,SB_198,SB_33,SB_44,SB_67,SB_72,SB_73,SB_74,SB_77,SB_85,SB_86

• Article ID: 5945
  Hykollari A, Balog CIA, Rendic D, Braulke W, Wilson IBH "Mass spectrometric analysis of neutral and anionic N-glycans from a Dictyostelium discoideum model for human congenital disorder of glycosylation CDG IL" - Journal of Proteome Research 12(3) (2013) 1173-1187
  

  The HL241 mutant strain of the cellular slime mold Dictyostelium discoideum is a potential model for human congenital disorder of glycosylation type IL (ALG9-CDG) and has been previously predicted to possess a lower degree of modification of its N-glycans with anionic moieties than the parental wild-type. In this study, we first showed that this strain has a premature stop codon in its alg9 mannosyltransferase gene compatible with the occurrence of truncated N-glycans. These were subject to an optimized analytical workflow, considering that the mass spectrometry of acidic glycans often presents challenges due to neutral loss and suppression effects. Therefore, the protein-bound N-glycans were first fractionated, after serial enzymatic release, by solid phase extraction. Then primarily single glycan species were isolated by mixed hydrophilic-interaction/anion-exchange or reversed-phase HPLC and analyzed using chemical and enzymatic treatments and MS/MS. We show that protein-linked N-glycans of the mutant are of reduced size as compared to those of wild-type AX3, but still contain core α1,3-fucose, intersecting N-acetylglucosamine, bisecting N-acetylglucosamine, methylphosphate, phosphate, and sulfate residues. We observe that a single N-glycan can carry up to four of these six possible modifications. Due to the improved analytical procedures, we reveal fuller details regarding the N-glycomic potential of this fascinating model organism.

  glycan, mass spectrometry, fucose, sulfate, Mannosyltransferase, N-glycans, Dictyostelium, methylphosphate

  NCBI PubMed ID: 23320427
  Publication DOI: 10.1021/pr300806b
  Journal NLM ID: 101128775
  Publisher: Washington, DC: American Chemical Society
  Correspondence: iain.wilson@boku.ac.at
  Institutions: Department für Chemie, Universität für Bodenkultur, A-1190 Wien, Austria, Biomolecular Mass Spectrometry Unit, Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands, Department of Biochemistry, Children's Hospital, University Medical Center Hamburg-Eppendorf, D-20246 Hamburg, Germany
  Methods: gel filtration, SDS-PAGE, sugar analysis, Western blotting, MALDI-TOF MS, enzymatic digestion, HF treatment, RT-PCR, cloning, RP-HPLC, RNA sequencing, HIAX-HPLC, MALDI-TOF MS/MS
  
   
  
  Dictyostelium discoideum HL241
  CSDB ID 4589 (all data & tools)

Show legend Show as text

Structure type: oligomer ; 1537 [M-H]-
Aglycon: 2-aminopyridine (PA)
Compound class: N-glycan
Contained glycoepitopes: IEDB_123886,IEDB_123888,IEDB_130701,IEDB_135813,IEDB_136045,IEDB_136104,IEDB_137340,IEDB_137485,IEDB_140116,IEDB_141793,IEDB_141807,IEDB_142489,IEDB_143632,IEDB_144562,IEDB_144983,IEDB_145669,IEDB_148493,IEDB_150092,IEDB_151531,IEDB_152206,IEDB_152214,IEDB_153212,IEDB_164174,IEDB_174333,IEDB_548907,IEDB_983930,SB_136,SB_196,SB_197,SB_198,SB_33,SB_44,SB_67,SB_72,SB_73,SB_74,SB_77,SB_85,SB_86

• Article ID: 5945
  Hykollari A, Balog CIA, Rendic D, Braulke W, Wilson IBH "Mass spectrometric analysis of neutral and anionic N-glycans from a Dictyostelium discoideum model for human congenital disorder of glycosylation CDG IL" - Journal of Proteome Research 12(3) (2013) 1173-1187
  

  The HL241 mutant strain of the cellular slime mold Dictyostelium discoideum is a potential model for human congenital disorder of glycosylation type IL (ALG9-CDG) and has been previously predicted to possess a lower degree of modification of its N-glycans with anionic moieties than the parental wild-type. In this study, we first showed that this strain has a premature stop codon in its alg9 mannosyltransferase gene compatible with the occurrence of truncated N-glycans. These were subject to an optimized analytical workflow, considering that the mass spectrometry of acidic glycans often presents challenges due to neutral loss and suppression effects. Therefore, the protein-bound N-glycans were first fractionated, after serial enzymatic release, by solid phase extraction. Then primarily single glycan species were isolated by mixed hydrophilic-interaction/anion-exchange or reversed-phase HPLC and analyzed using chemical and enzymatic treatments and MS/MS. We show that protein-linked N-glycans of the mutant are of reduced size as compared to those of wild-type AX3, but still contain core α1,3-fucose, intersecting N-acetylglucosamine, bisecting N-acetylglucosamine, methylphosphate, phosphate, and sulfate residues. We observe that a single N-glycan can carry up to four of these six possible modifications. Due to the improved analytical procedures, we reveal fuller details regarding the N-glycomic potential of this fascinating model organism.

  glycan, mass spectrometry, fucose, sulfate, Mannosyltransferase, N-glycans, Dictyostelium, methylphosphate

  NCBI PubMed ID: 23320427
  Publication DOI: 10.1021/pr300806b
  Journal NLM ID: 101128775
  Publisher: Washington, DC: American Chemical Society
  Correspondence: iain.wilson@boku.ac.at
  Institutions: Department für Chemie, Universität für Bodenkultur, A-1190 Wien, Austria, Biomolecular Mass Spectrometry Unit, Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands, Department of Biochemistry, Children's Hospital, University Medical Center Hamburg-Eppendorf, D-20246 Hamburg, Germany
  Methods: gel filtration, SDS-PAGE, sugar analysis, Western blotting, MALDI-TOF MS, enzymatic digestion, HF treatment, RT-PCR, cloning, RP-HPLC, RNA sequencing, HIAX-HPLC, MALDI-TOF MS/MS
  
   
  
  Dictyostelium discoideum HL241
  CSDB ID 4590 (all data & tools)

Show legend Show as text

Structure type: oligomer ; 1631 [M-H]-
Aglycon: 2-aminopyridine (PA)
Compound class: N-glycan
Contained glycoepitopes: IEDB_123886,IEDB_123888,IEDB_130701,IEDB_135813,IEDB_136045,IEDB_136104,IEDB_137340,IEDB_137485,IEDB_140116,IEDB_141793,IEDB_141807,IEDB_142489,IEDB_143632,IEDB_144562,IEDB_144983,IEDB_145669,IEDB_148493,IEDB_150092,IEDB_151531,IEDB_152206,IEDB_152214,IEDB_153212,IEDB_164174,IEDB_174333,IEDB_474450,IEDB_548907,IEDB_983930,SB_136,SB_196,SB_197,SB_198,SB_33,SB_44,SB_67,SB_72,SB_73,SB_74,SB_77,SB_85,SB_86

• Article ID: 5945
  Hykollari A, Balog CIA, Rendic D, Braulke W, Wilson IBH "Mass spectrometric analysis of neutral and anionic N-glycans from a Dictyostelium discoideum model for human congenital disorder of glycosylation CDG IL" - Journal of Proteome Research 12(3) (2013) 1173-1187
  

  The HL241 mutant strain of the cellular slime mold Dictyostelium discoideum is a potential model for human congenital disorder of glycosylation type IL (ALG9-CDG) and has been previously predicted to possess a lower degree of modification of its N-glycans with anionic moieties than the parental wild-type. In this study, we first showed that this strain has a premature stop codon in its alg9 mannosyltransferase gene compatible with the occurrence of truncated N-glycans. These were subject to an optimized analytical workflow, considering that the mass spectrometry of acidic glycans often presents challenges due to neutral loss and suppression effects. Therefore, the protein-bound N-glycans were first fractionated, after serial enzymatic release, by solid phase extraction. Then primarily single glycan species were isolated by mixed hydrophilic-interaction/anion-exchange or reversed-phase HPLC and analyzed using chemical and enzymatic treatments and MS/MS. We show that protein-linked N-glycans of the mutant are of reduced size as compared to those of wild-type AX3, but still contain core α1,3-fucose, intersecting N-acetylglucosamine, bisecting N-acetylglucosamine, methylphosphate, phosphate, and sulfate residues. We observe that a single N-glycan can carry up to four of these six possible modifications. Due to the improved analytical procedures, we reveal fuller details regarding the N-glycomic potential of this fascinating model organism.

  glycan, mass spectrometry, fucose, sulfate, Mannosyltransferase, N-glycans, Dictyostelium, methylphosphate

  NCBI PubMed ID: 23320427
  Publication DOI: 10.1021/pr300806b
  Journal NLM ID: 101128775
  Publisher: Washington, DC: American Chemical Society
  Correspondence: iain.wilson@boku.ac.at
  Institutions: Department für Chemie, Universität für Bodenkultur, A-1190 Wien, Austria, Biomolecular Mass Spectrometry Unit, Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands, Department of Biochemistry, Children's Hospital, University Medical Center Hamburg-Eppendorf, D-20246 Hamburg, Germany
  Methods: gel filtration, SDS-PAGE, sugar analysis, Western blotting, MALDI-TOF MS, enzymatic digestion, HF treatment, RT-PCR, cloning, RP-HPLC, RNA sequencing, HIAX-HPLC, MALDI-TOF MS/MS
  
   
  
  Dictyostelium discoideum HL241
  CSDB ID 4591 (all data & tools)

Show legend Show as text

Structure type: oligomer ; 1865 [M+H]+
Aglycon: 2-aminopyridine (PA)
Compound class: N-glycan
Contained glycoepitopes: IEDB_123886,IEDB_123888,IEDB_130701,IEDB_135813,IEDB_136045,IEDB_136104,IEDB_137340,IEDB_137485,IEDB_140116,IEDB_141793,IEDB_141807,IEDB_142489,IEDB_143632,IEDB_144562,IEDB_144983,IEDB_145669,IEDB_148493,IEDB_150092,IEDB_151531,IEDB_152206,IEDB_152214,IEDB_153212,IEDB_164174,IEDB_174333,IEDB_548907,IEDB_983930,SB_136,SB_196,SB_197,SB_198,SB_33,SB_44,SB_67,SB_72,SB_73,SB_74,SB_77,SB_85,SB_86

• Article ID: 5945
  Hykollari A, Balog CIA, Rendic D, Braulke W, Wilson IBH "Mass spectrometric analysis of neutral and anionic N-glycans from a Dictyostelium discoideum model for human congenital disorder of glycosylation CDG IL" - Journal of Proteome Research 12(3) (2013) 1173-1187
  

  The HL241 mutant strain of the cellular slime mold Dictyostelium discoideum is a potential model for human congenital disorder of glycosylation type IL (ALG9-CDG) and has been previously predicted to possess a lower degree of modification of its N-glycans with anionic moieties than the parental wild-type. In this study, we first showed that this strain has a premature stop codon in its alg9 mannosyltransferase gene compatible with the occurrence of truncated N-glycans. These were subject to an optimized analytical workflow, considering that the mass spectrometry of acidic glycans often presents challenges due to neutral loss and suppression effects. Therefore, the protein-bound N-glycans were first fractionated, after serial enzymatic release, by solid phase extraction. Then primarily single glycan species were isolated by mixed hydrophilic-interaction/anion-exchange or reversed-phase HPLC and analyzed using chemical and enzymatic treatments and MS/MS. We show that protein-linked N-glycans of the mutant are of reduced size as compared to those of wild-type AX3, but still contain core α1,3-fucose, intersecting N-acetylglucosamine, bisecting N-acetylglucosamine, methylphosphate, phosphate, and sulfate residues. We observe that a single N-glycan can carry up to four of these six possible modifications. Due to the improved analytical procedures, we reveal fuller details regarding the N-glycomic potential of this fascinating model organism.

  glycan, mass spectrometry, fucose, sulfate, Mannosyltransferase, N-glycans, Dictyostelium, methylphosphate

  NCBI PubMed ID: 23320427
  Publication DOI: 10.1021/pr300806b
  Journal NLM ID: 101128775
  Publisher: Washington, DC: American Chemical Society
  Correspondence: iain.wilson@boku.ac.at
  Institutions: Department für Chemie, Universität für Bodenkultur, A-1190 Wien, Austria, Biomolecular Mass Spectrometry Unit, Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands, Department of Biochemistry, Children's Hospital, University Medical Center Hamburg-Eppendorf, D-20246 Hamburg, Germany
  Methods: gel filtration, SDS-PAGE, sugar analysis, Western blotting, MALDI-TOF MS, enzymatic digestion, HF treatment, RT-PCR, cloning, RP-HPLC, RNA sequencing, HIAX-HPLC, MALDI-TOF MS/MS
  
   
  
  Dictyostelium discoideum HL241
  CSDB ID 4594 (all data & tools)

Show legend Show as text

Structure type: oligomer ; 2351 [M+H]+
Aglycon: 2-aminopyridine (PA)
Compound class: N-glycan
Contained glycoepitopes: IEDB_123886,IEDB_123888,IEDB_130701,IEDB_135813,IEDB_136045,IEDB_136104,IEDB_137340,IEDB_137485,IEDB_140116,IEDB_141793,IEDB_141807,IEDB_141828,IEDB_141829,IEDB_141831,IEDB_142489,IEDB_143632,IEDB_144562,IEDB_144983,IEDB_145669,IEDB_148493,IEDB_150092,IEDB_151079,IEDB_151531,IEDB_152206,IEDB_152214,IEDB_153212,IEDB_153220,IEDB_164174,IEDB_174333,IEDB_187201,IEDB_429156,IEDB_548907,IEDB_857734,IEDB_983930,SB_136,SB_191,SB_196,SB_197,SB_198,SB_33,SB_44,SB_53,SB_67,SB_72,SB_73,SB_74,SB_77,SB_85,SB_86

• Article ID: 5945
  Hykollari A, Balog CIA, Rendic D, Braulke W, Wilson IBH "Mass spectrometric analysis of neutral and anionic N-glycans from a Dictyostelium discoideum model for human congenital disorder of glycosylation CDG IL" - Journal of Proteome Research 12(3) (2013) 1173-1187
  

  The HL241 mutant strain of the cellular slime mold Dictyostelium discoideum is a potential model for human congenital disorder of glycosylation type IL (ALG9-CDG) and has been previously predicted to possess a lower degree of modification of its N-glycans with anionic moieties than the parental wild-type. In this study, we first showed that this strain has a premature stop codon in its alg9 mannosyltransferase gene compatible with the occurrence of truncated N-glycans. These were subject to an optimized analytical workflow, considering that the mass spectrometry of acidic glycans often presents challenges due to neutral loss and suppression effects. Therefore, the protein-bound N-glycans were first fractionated, after serial enzymatic release, by solid phase extraction. Then primarily single glycan species were isolated by mixed hydrophilic-interaction/anion-exchange or reversed-phase HPLC and analyzed using chemical and enzymatic treatments and MS/MS. We show that protein-linked N-glycans of the mutant are of reduced size as compared to those of wild-type AX3, but still contain core α1,3-fucose, intersecting N-acetylglucosamine, bisecting N-acetylglucosamine, methylphosphate, phosphate, and sulfate residues. We observe that a single N-glycan can carry up to four of these six possible modifications. Due to the improved analytical procedures, we reveal fuller details regarding the N-glycomic potential of this fascinating model organism.

  glycan, mass spectrometry, fucose, sulfate, Mannosyltransferase, N-glycans, Dictyostelium, methylphosphate

  NCBI PubMed ID: 23320427
  Publication DOI: 10.1021/pr300806b
  Journal NLM ID: 101128775
  Publisher: Washington, DC: American Chemical Society
  Correspondence: iain.wilson@boku.ac.at
  Institutions: Department für Chemie, Universität für Bodenkultur, A-1190 Wien, Austria, Biomolecular Mass Spectrometry Unit, Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands, Department of Biochemistry, Children's Hospital, University Medical Center Hamburg-Eppendorf, D-20246 Hamburg, Germany
  Methods: gel filtration, SDS-PAGE, sugar analysis, Western blotting, MALDI-TOF MS, enzymatic digestion, HF treatment, RT-PCR, cloning, RP-HPLC, RNA sequencing, HIAX-HPLC, MALDI-TOF MS/MS
  
   
  
  Dictyostelium discoideum AX3
  CSDB ID 8141 (all data & tools)

Show legend Show as text

Structure type: oligomer ; 2429 [M-H]-
Aglycon: 2-aminopyridine (PA)
Compound class: N-glycan
Contained glycoepitopes: IEDB_123886,IEDB_123888,IEDB_130701,IEDB_135813,IEDB_136045,IEDB_136104,IEDB_137340,IEDB_137485,IEDB_140116,IEDB_141793,IEDB_141807,IEDB_141828,IEDB_141829,IEDB_141831,IEDB_142489,IEDB_143632,IEDB_144562,IEDB_144983,IEDB_145669,IEDB_148493,IEDB_150092,IEDB_151079,IEDB_151531,IEDB_152206,IEDB_152214,IEDB_153212,IEDB_153220,IEDB_164174,IEDB_174333,IEDB_187201,IEDB_429156,IEDB_548907,IEDB_857734,IEDB_983930,SB_136,SB_191,SB_196,SB_197,SB_198,SB_33,SB_44,SB_53,SB_67,SB_72,SB_73,SB_74,SB_77,SB_85,SB_86

• Article ID: 5945
  Hykollari A, Balog CIA, Rendic D, Braulke W, Wilson IBH "Mass spectrometric analysis of neutral and anionic N-glycans from a Dictyostelium discoideum model for human congenital disorder of glycosylation CDG IL" - Journal of Proteome Research 12(3) (2013) 1173-1187
  

  The HL241 mutant strain of the cellular slime mold Dictyostelium discoideum is a potential model for human congenital disorder of glycosylation type IL (ALG9-CDG) and has been previously predicted to possess a lower degree of modification of its N-glycans with anionic moieties than the parental wild-type. In this study, we first showed that this strain has a premature stop codon in its alg9 mannosyltransferase gene compatible with the occurrence of truncated N-glycans. These were subject to an optimized analytical workflow, considering that the mass spectrometry of acidic glycans often presents challenges due to neutral loss and suppression effects. Therefore, the protein-bound N-glycans were first fractionated, after serial enzymatic release, by solid phase extraction. Then primarily single glycan species were isolated by mixed hydrophilic-interaction/anion-exchange or reversed-phase HPLC and analyzed using chemical and enzymatic treatments and MS/MS. We show that protein-linked N-glycans of the mutant are of reduced size as compared to those of wild-type AX3, but still contain core α1,3-fucose, intersecting N-acetylglucosamine, bisecting N-acetylglucosamine, methylphosphate, phosphate, and sulfate residues. We observe that a single N-glycan can carry up to four of these six possible modifications. Due to the improved analytical procedures, we reveal fuller details regarding the N-glycomic potential of this fascinating model organism.

  glycan, mass spectrometry, fucose, sulfate, Mannosyltransferase, N-glycans, Dictyostelium, methylphosphate

  NCBI PubMed ID: 23320427
  Publication DOI: 10.1021/pr300806b
  Journal NLM ID: 101128775
  Publisher: Washington, DC: American Chemical Society
  Correspondence: iain.wilson@boku.ac.at
  Institutions: Department für Chemie, Universität für Bodenkultur, A-1190 Wien, Austria, Biomolecular Mass Spectrometry Unit, Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands, Department of Biochemistry, Children's Hospital, University Medical Center Hamburg-Eppendorf, D-20246 Hamburg, Germany
  Methods: gel filtration, SDS-PAGE, sugar analysis, Western blotting, MALDI-TOF MS, enzymatic digestion, HF treatment, RT-PCR, cloning, RP-HPLC, RNA sequencing, HIAX-HPLC, MALDI-TOF MS/MS
  
   
  
  Dictyostelium discoideum AX3
  CSDB ID 8142 (all data & tools)

Show legend Show as text

Structure type: oligomer
Compound class: N-glycan
Contained glycoepitopes: IEDB_123886,IEDB_123888,IEDB_130701,IEDB_135813,IEDB_136045,IEDB_136104,IEDB_137340,IEDB_137485,IEDB_140116,IEDB_141793,IEDB_141807,IEDB_141828,IEDB_141829,IEDB_141831,IEDB_142489,IEDB_143632,IEDB_144562,IEDB_144983,IEDB_145669,IEDB_148493,IEDB_150092,IEDB_151079,IEDB_151531,IEDB_152206,IEDB_152214,IEDB_153212,IEDB_153220,IEDB_164174,IEDB_174333,IEDB_187201,IEDB_429156,IEDB_474450,IEDB_548907,IEDB_857734,IEDB_983930,SB_136,SB_191,SB_196,SB_197,SB_198,SB_33,SB_44,SB_53,SB_67,SB_72,SB_73,SB_74,SB_77,SB_85,SB_86

• Article ID: 5990
  Paschinger K, Wilson IBH "Analysis of zwitterionic and anionic N-linked glycans from invertebrates and protists by mass spectrometry" - Glycoconjugate Journal 33(3) (2016) 273-283
  

  Glycomic analyses over the years have revealed that non-vertebrate eukaryotes express oligosaccharides with inorganic and zwitterionic modifications which are either occurring in different contexts as compared to, or are absent from, mammals. Examples of anionic N-glycans (carrying sulphate or phosphate) are known from amoebae, fungi, molluscs and insects, while zwitterionic modifications by phosphorylcholine, phosphoethanolamine and aminoethylphosphonate occur on N-, O- and lipid-linked glycans from trichomonads, annelids, fungi, molluscs, insects, cestodes and nematodes. For detection of zwitterionic and anionic glycans, mass spectrometry has been a key method, but their ionic character affects the preparation and purification; therefore, as part of a glycomic strategy, the possibility of their presence must be considered in advance. On the other hand, their ionisation and fragmentation in positive and negative ion mode mass spectrometry as well as specific chemical or enzymatic treatments can prove diagnostic to their analysis. In our laboratory, we combine solid-phase extraction, reversed and normal phase HPLC, MALDI-TOF MS, exoglycosidase digests and hydrofluoric acid treatment to reveal N-glycans modified with anionic and zwitterionic moieties in a wide range of organisms. It is to be anticipated that, as more species are glycomically analysed, zwitterionic and anionic modifications of N-glycans will prove rather widespread. This knowledge is - in the longer term - then the basis for understanding the function of this cornucopia of glycan modifications.

  phosphate, mass spectrometry, phosphoethanolamine, phosphorylcholine, Glycomics, Aminoethylphosphonate, Glucuronate, Sulphate

  NCBI PubMed ID: 26899268
  Publication DOI: 10.1007/s10719-016-9650-x
  Journal NLM ID: 8603310
  Publisher: Kluwer Academic Publishers
  Correspondence: iain.wilson@boku.ac.at
  Institutions: Department für Chemie, Universität für Bodenkultur, 1190, Wien, Austria
  
   
  
  Dictyostelium discoideum
  CSDB ID 8298 (all data & tools)

Show legend Show as text

Structure type: oligomer
Compound class: N-glycan
Contained glycoepitopes: IEDB_123886,IEDB_123888,IEDB_130701,IEDB_135813,IEDB_136045,IEDB_136104,IEDB_137340,IEDB_137485,IEDB_140116,IEDB_141793,IEDB_141807,IEDB_141828,IEDB_141829,IEDB_141830,IEDB_141831,IEDB_142489,IEDB_143632,IEDB_144562,IEDB_144983,IEDB_145669,IEDB_148493,IEDB_150092,IEDB_151079,IEDB_151531,IEDB_152206,IEDB_152214,IEDB_153212,IEDB_153220,IEDB_164174,IEDB_174333,IEDB_187201,IEDB_429156,IEDB_540671,IEDB_548907,IEDB_857734,IEDB_983930,SB_136,SB_191,SB_196,SB_197,SB_198,SB_33,SB_44,SB_53,SB_67,SB_72,SB_73,SB_74,SB_77,SB_85,SB_86

• Article ID: 5996
  Schiller B, Hykollari A, Yan S, Paschinger K, Wilson IBH "Complicated N-linked glycans in simple organisms" - Biological Chemistry 393(8) (2012) 661-673
  

  Although countless genomes have now been sequenced, the glycomes of the vast majority of eukaryotes still present a series of unmapped frontiers. However, strides are being made in a few groups of invertebrate and unicellular organisms as regards their N-glycans and N-glycosylation pathways. Thereby, the traditional classification of glycan structures inevitably approaches its boundaries. Indeed, the glycomes of these organisms are rich in surprises including a multitude of modifications of the core regions of N-glycans and unusual antennae. From the actually rather limited glycomic information we have, it is nevertheless obvious that the biotechnological, developmental and immunological relevance of these modifications, especially in insect cell lines, model organisms and parasites means that deciphering unusual glycomes is of more than just academic interest.

  N-linked oligosaccharides, protozoa, insects, molluscs, nematodes, trematodes

  NCBI PubMed ID: 22944671
  Publication DOI: 10.1515/hsz-2012-0150
  Journal NLM ID: 9700112
  Publisher: Berlin: Walter De Gruyter
  Correspondence: iain.wilson@boku.ac.at
  Institutions: Department für Chemie, Universität für Bodenkultur, 1190 Wien, Austria
  
   
  
  Dictyostelium discoideum
  CSDB ID 8369 (all data & tools)

Show legend Show as text

Structure type: oligomer
Compound class: N-glycan, glucomannan, oligosaccharide
Contained glycoepitopes: IEDB_123886,IEDB_123888,IEDB_130701,IEDB_135813,IEDB_136045,IEDB_137340,IEDB_137485,IEDB_141793,IEDB_141807,IEDB_142489,IEDB_144562,IEDB_144983,IEDB_145669,IEDB_148493,IEDB_150092,IEDB_151531,IEDB_152206,IEDB_152214,IEDB_153212,IEDB_174333,IEDB_548907,IEDB_983930,SB_197,SB_198,SB_33,SB_44,SB_67,SB_72,SB_73,SB_74,SB_85,SB_86

• Article ID: 6014
  Tjondro HC, Loke I, Chatterjee S, Thaysen-Andersen M "Human protein paucimannosylation: cues from the eukaryotic kingdoms" - Biological Reviews of the Cambridge Philosophical Society 94(6) (2019) 2068-2100
  

  Paucimannosidic proteins (PMPs) are bioactive glycoproteins carrying truncated α- or β-mannosyl-terminating asparagine (N)-linked glycans widely reported across the eukaryotic domain. Our understanding of human PMPs remains limited, despite findings documenting their existence and association with human disease glycobiology. This review comprehensively surveys the structures, biosynthetic routes and functions of PMPs across the eukaryotic kingdoms with the aim of synthesising an improved understanding on the role of protein paucimannosylation in human health and diseases. Convincing biochemical, glycoanalytical and biological data detail a vast structural heterogeneity and fascinating tissue- and subcellular-specific expression of PMPs within invertebrates and plants, often comprising multi-α1,3/6-fucosylation and β1,2-xylosylation amongst other glycan modifications and non-glycan substitutions e.g. O-methylation. Vertebrates and protists express less-heterogeneous PMPs typically only comprising variable core fucosylation of bi- and trimannosylchitobiose core glycans. In particular, the Manα1,6Manβ1,4GlcNAc(α1,6Fuc)β1,4GlcNAcβAsn glycan (M2F) decorates various human neutrophil proteins reportedly displaying bioactivity and structural integrity demonstrating that they are not degradation products. Less-truncated paucimannosidic glycans (e.g. M3F) are characteristic glycosylation features of proteins expressed by human cancer and stem cells. Concertedly, these observations suggest the involvement of human PMPs in processes related to innate immunity, tumorigenesis and cellular differentiation. The absence of human PMPs in diverse bodily fluids studied under many (patho)physiological conditions suggests extravascular residence and points to localised functions of PMPs in peripheral tissues. Absence of PMPs in Fungi indicates that paucimannosylation is common, but not universally conserved, in eukaryotes. Relative to human PMPs, the expression of PMPs in plants, invertebrates and protists is more tissue-wide and constitutive yet, similar to their human counterparts, PMP expression remains regulated by the physiology of the producing organism and PMPs evidently serve essential functions in development, cell-cell communication and host-pathogen/symbiont interactions. In most PMP-producing organisms, including humans, the N-acetyl-β-hexosaminidase isoenzymes and linkage-specific α-mannosidases are glycoside hydrolases critical for generating PMPs via N-acetylglucosaminyltransferase I (GnT-I)-dependent and GnT-I-independent truncation pathways. However, the identity and structure of many species-specific PMPs in eukaryotes, their biosynthetic routes, strong tissue- and development-specific expression, and diverse functions are still elusive. Deep exploration of these PMP features involving, for example, the characterisation of endogenous PMP-recognising lectins across a variety of healthy and N-acetyl-β-hexosaminidase-deficient human tissue types and identification of microbial adhesins reactive to human PMPs, are amongst the many tasks required for enhanced insight into the glycobiology of human PMPs. In conclusion, the literature supports the notion that PMPs are significant, yet still heavily under-studied biomolecules in human glycobiology that serve essential functions and create structural heterogeneity not dissimilar to other human N-glycoprotein types. Human PMPs should therefore be recognised as bioactive glycoproteins that are distinctly different from the canonical N-glycoprotein classes and which warrant a more dedicated focus in glycobiological research.

  innate immunity, N-glycosylation, glycobiology, eukaryote, N-acetyl-β-hexosaminidase, N -acetylglucosaminyltransferase I, paucimannosidic protein, paucimannosylation

  NCBI PubMed ID: 31410980
  Publication DOI: 10.1111/brv.12548
  Journal NLM ID: 0414576
  Publisher: London, Cambridge University Press
  Correspondence: morten.andersen@mq.edu.au
  Institutions: Department of Molecular Sciences, Macquarie University, Sydney, New South Wales, 2109, Australia, Department of Biological Sciences, National University of Singapore, Singapore 117543, Singapore
  
   
  
  Dictyostelium discoideum
  CSDB ID 9477 (all data & tools)
• Article ID: 11783
  Choi KH, Laursen RA "Amino-acid sequence and glycan structures of cysteine proteases with proline specificity from ginger rhizome Zingiber officinale" - European Journal of Biochemistry 267(5) (2000) 1516-1526
  

  The ginger proteases (GP-I and GP-II), isolated from the ginger rhizome Zingiber officinale, have an unusual substrate specificity preference for cleaving peptides with a proline residue at the P2 position. The complete amino-acid sequence of GP-II, a glycoprotein containing 221 amino acids, and about 98% that of GP-I have been determined. Both proteases, which are 82% similar, have cysteine residues at positions 27 and histidines at position 161, corresponding to the essential cysteine-histidine diads found in the papain family of cysteine proteases, and six corresponding cysteine residues that form the three invariant disulfide linkages seen in this family of proteins. The sequence homology with other members (papain, bromelain, actinidin, protease omega, etc.) of this family is approximately 50%. GP-II has two predicted glycosylation sites at Asn99 and Asn156. Analyisis by electrospray and collision-induced dissociation MS showed that both sites were occupied by the glycans (Man)3(Xyl)1(Fuc)1(GlcNAc)2 and (Man)3(Xyl)1(Fuc)1(GlcNAc)3, in a ratio of approximately 7 : 1. Both glycans are xylose containing biantennary complex types that share the common core structural unit, Man1→6(Man1→3)(Xyl1→2)Man1→4GlcNAc1→4(Fuc1→3)GlcNAc for the major form, with an additional N-acetylglucosamine residue being linked, in the minor form, to one of the terminal mannose units of the core structure.

  mass spectrometry, glycoprotein, amino-acid sequence, cysteine protease, ginger, glycosylation site, proline peptidase, proteinase

  NCBI PubMed ID: 10691991
  Publication DOI: 10.1046/j.1432-1327.2000.01152.x
  Journal NLM ID: 0107600
  Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
  Correspondence: laursen@bu.edu
  Institutions: Department of Chemistry, Boston University, Boston, USA
  Methods: methylation, chemical analysis, ESI-MS, acid hydrolysis, GLC, MALDI-TOF MS, HPLC, UV, enzymatic digestion, extraction, CID-MS, acetylation, reduction, column chromatography, RP-HPLC, dialysis, enzymatic assay, precipitation, derivatization, centrifugation, amino acid sequence analysis
  
   
  
  Ananas comosus
  CSDB ID 65148 (all data & tools)
• Article ID: 12256
  Olczak M, Watorek W "Structural analysis of N-glycans from yellow lupin (Lupinus luteus) seed diphosphonucleotide phosphatase/phosphodiesterase" - Biochimica et Biophysica Acta 1523(2-3) (2000) 236-245
  

  N-linked oligosaccharide chains released by hydrazinolysis from yellow lupin seed diphosphonucleotide phosphatase/phosphodiesterase were fluorescence labeled and separated by high performance liquid chromatography (GlycoSep N and GlycoSep H columns). Exoglycosidase sequencing elucidated the structures of 24 separated N-glycans. Thirty percent of isolated glycans were found to be of high-mannose type (three to eight mannosyl residues), 42% were complex type and 26% belonged to paucimannosidic type. Among complex type glycans, structures with Lewis(a) epitope were identified. It is very unusual to find all types of plant N-glycans in one protein. Possible reasons for such a broad spectrum of N-glycan structures are discussed.

  N-glycan, exoglycosidase sequencing, diphosphonucleotide phosphatase/phosphodiesterase, yellow lupin

  NCBI PubMed ID: 11042390
  Publication DOI: 10.1016/s0304-4165(00)00128-8
  Journal NLM ID: 0217513
  Publisher: Elsevier
  Correspondence: watorek@bf.uni.wroc.pl
  Institutions: Institute of Biochemistry and Molecular Biology, Wroclaw University, Wroclaw, Poland
  Methods: HPLC, enzymatic digestion, extraction, fluorescence labeling, spectrometry
  
   
  
  Lupinus luteus
  CSDB ID 67228 (all data & tools)

Show legend Show as text

Structure type: oligomer
Compound class: N-glycan
Contained glycoepitopes: IEDB_114701,IEDB_115005,IEDB_116644,IEDB_122244,IEDB_123886,IEDB_123887,IEDB_123888,IEDB_130701,IEDB_135813,IEDB_136045,IEDB_137340,IEDB_137485,IEDB_141793,IEDB_141807,IEDB_142489,IEDB_144562,IEDB_144983,IEDB_145668,IEDB_145669,IEDB_148491,IEDB_148492,IEDB_148493,IEDB_150092,IEDB_151531,IEDB_152206,IEDB_152214,IEDB_153212,IEDB_167188,IEDB_174332,IEDB_174333,IEDB_548907,IEDB_983930,SB_197,SB_198,SB_33,SB_44,SB_67,SB_72,SB_73,SB_74,SB_85,SB_86

• Article ID: 9546
  Takahashi N, Hotta T, Ishihara H, Mori M, Tejima S, Bligny R, Akazawa T, Endo S, Arata Y "Xylose-containing common structural unit in N-linked oligosaccharides of laccase from sycamore cells" - Biochemistry 25(2) (1986) 388-395
  

  The structures of asparagine-linked oligosaccharides of laccase excreted by sycamore (Acer pseudoplatanus L.) cells are reported. Peptic glycopeptides obtained from the laccase were treated with N-oligosaccharide glycopeptidase (EC 3.5.1.52) to release the oligosaccharide moieties. The oligosaccharides thus obtained were fractionated into six components by gel filtration, thin-layer chromatography, and high-performance liquid chromatography. The structures of the isolated oligosaccharides were determined by sugar analysis, exoglycosidase digestion, and methylation analysis in combination with high-resolution proton nuclear magnetic resonance spectroscopy. It was found that (1) the six oligosaccharides are a series of compounds of xylose-containing biantennary complex types that share as the core a common structural unit, i.e., Xyl-β(l-2)[Man-α(l-6)]Man-β(l-4)GlcNAc-β(l-4)[Fuc-α(1-3)]GlcNAc, and (2) mannose, N-acetylglucosamine, galactose, and fucose residues are additionally linked to the core as the outer chain moieties.

  Publication DOI: 10.1021/bi00350a018
  Journal NLM ID: 0370623
  Publisher: American Chemical Society
  Institutions: Department of Hygienic Chemistry, Faculty of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan, Department of Biochemistry, Nagoya City University Medical School, Nagoya, Japan, Centre National de la Recherche Scientifique, CENG, Grenoble, France, Research Institute for Biological Regulation School of Agriculture, Nagoya University, Nagoya, Japan, Department of Biophysics and Biochemistry, Faculty of Science, University of Tokyo, Tokyo, Japan
  Methods: 1H NMR, TLC, acid hydrolysis, HPLC, enzymatic digestion, methylation analysis
  
   
  
  Acer pseudoplatanus
  CSDB ID 107206 (all data & tools)
• Article ID: 9930
  Priem B, Solokwan J, Wieruszeski JM, Strecker G, Nazih H, Morvan H "Isolation and characterization of free glycans of the oligomannoside type from the extracellular medium of a plant cell suspension" - Glycoconjugate Journal 7 (1990) 121-132
  

  The oligosaccharides Man5GlcNAc and Man3(Xyl)GlcNAc(Fuc)GlcNAc presumed to originate fromN-glycosyl proteins have been purified from an extracellular medium (concentration: 2–5 mg/l of 14 day cultures) of white campion (Silene alba) suspension culture. Their primary structures have been determined by1H-400-MHz NMR spectroscopy and FAB-MS spectrometry. They are probably the result of an autophagic process including protein catabolism due to sucrose starvation. Additional identification of digalactosylglycerol (galactolipid breakdown) argues for this hypothesis.

  oligosaccharide, glycan, glycoconjugate, white campion

  Journal NLM ID: 8603310
  Publisher: Kluwer Academic Publishers
  Institutions: Equipe Polysaccharides Pariétaux des Végétaux, Université des Sciences et Techniques de Lille Flandres-Artois, Villeneuve d'Ascq Cédex, France, Laboratoire de Chimie-Biologique, Université des Sciences et Techniques de Lille Flandres-Artois, Villeneuve d'Ascq Cédex, France
  Methods: gel filtration, 1H NMR, FAB-MS, TLC, GLC, permethylation, PE
  
   
  
  Silene alba
  CSDB ID 128195 (all data & tools)
• Article ID: 10051
  Lhernould S, Karamanos Y, Bourgerie S, Strecker G, Julien R, Morvan H "Peptide-N4-(N-acetylglucosaminyl)asparagine amidase (PNGase) activity could explain the occurrence of extracellular xylomannosides in a plant cell suspension" - Glycoconjugate Journal 9 (1992) 191-197
  

  We have previously isolated mannoside and xylomannoside oligosaccharides with one or two terminal reducing N-acetylglucosamine residues from the extracellular medium of white campion (Silene alba) suspension culture. We have now demonstrated the presence of peptide-N4-(N-acetylglucosaminyl)asparagine amidase (PNGase) activity in cell extracts as well in the culture medium that could explain the production of those compounds. An additional xylomannoside, (GlcNAc)Man3(Xyl)GlcNAc(Fuc)GlcNAc, was characterized, and 1H- and 13C-NMR assignments for the oligosaccharide Man3(Xyl)GlcNAc(Fuc)GlcNAc were obtained using homonuclear and heteronuclear spectroscopy (COSY).

  white campion, xylomannoside, PNGase

  NCBI PubMed ID: 1422139
  Journal NLM ID: 8603310
  Publisher: Kluwer Academic Publishers
  Institutions: Laboratoire de Biologie Cellulaire Vegetale et Valorisation des Especes Ligneuses, Université de Limoges, Limoges, France
  Methods: 1H NMR, enzymatic digestion, PPC, 13C
  
   
  
  Silene alba
  CSDB ID 128413 (all data & tools)
• Article ID: 10301
  Ashford D, Dwek RA, Welply JK, Amatayakul S, Homans SW, Lis H, Taylor GN, Sharon N, Rademacher TW "The β-1-2-D-xylose and α-1-3-L-fucose substituted N-linked oligosaccharides from Erythrina crista-galli lectin. Isolation, characterization and comparison with other legume lectins" - European Journal of Biochemistry 166 (1987) 311-320
  

  The carbohydrate moieties of Erythrina cristagalli lectin were released as oligosaccharides by hydrazinolysis, followed by N-acetylation and reduction with NaB3H4. Fractionation of the tritium-labelled oligosaccharide mixture by Bio-Gel P-4 column chromatography and high-voltage borate electrophoresis revealed that it is composed of five neutral oligosaccharides. Structural studies by sequential exoglycosidase digestion in combination with methylation analysis and two-dimensional 1H-NMR showed that the major component was the fucose-containing heptasaccharide Man α 3(Man α 6)(Xyl β 2)Man β 4GlcNAc β 4(Fuc α 3)GlcNAcol. This is the first report of such a structure in plant lectins. Small amounts of the corresponding afucosyl hexasaccharide were also identified, as well as three other minor components. The structure of the heptasaccharide shows the twin characteristics of a newly established family of N-linked glycans, found to date only in plants. The characteristics are substitution of the common pentasaccharide core [Man α 3(Man α 6)Man β 4GlcNAc β 4GlcNAc] by a D-xylose residue linked β1----2 to the β-mannosyl residue and an L-fucose residue linked α1----3 to the reducing terminal N-acetylglucosamine residue. The oligosaccharide heterogeneity pattern for Erythrina cristagalli lectin was also found for the lectins from four other Erythrina species and the lectins of two other legumes, Sophora japonica and Lonchocarpus capassa.

  NCBI PubMed ID: 3609010
  Publication DOI: 10.1111/j.1432-1033.1987.tb13516.x
  Journal NLM ID: 0107600
  Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
  Institutions: Department of Biochemistry, University of Oxford, Oxford, UK, Department of Biophysics, The Weizmann Institute of Science, Rehovot, Israel
  Methods: gel filtration, 1H NMR, GC-MS, enzymatic digestion, methylation analysis, reduction, PC
  
   
  
  Erythrina crista-galli
  CSDB ID 123040 (all data & tools)
• Article ID: 10683
  Kimura Y, Hase S, Kobayashi Y, Kyogoku Y, Ikenaka T, Funatsu G "Structures of sugar chains of Ricinus communis agglutinin" - Biochimica et Biophysica Acta 966 (1988) 248-256
  

  The structures of sugar chains from Ricinus communis agglutinin were determined. Four glycopeptides were isolated from the lectin according to a published method (Kimura, Y. and Funatsu, G. (1988) Agric. Biol. Chem. 52, in press), and sugar chains of each glycopeptide were liberated by hydrazinolysis. Free amino groups were N-acetylated and the reducing-end residues were coupled with 2-aminopyridine. The resulting pyridylamino derivatives of sugar chains were purified by gel filtration and reversed-phase HPLC. The structures of thus-purified PA-sugar chains were determined by a combination of component analysis, stepwise exoglycosidase digestions, partial acetolysis, and 500 MHz 1H-NMR spectroscopy. These results indicate that R. communis agglutinin contains the sugar chains shown on page 249.

  lectin, glycoprotein, Sugar chain structure, Pyridylamino derivative, Sugar chain processing, Ricinus communis agglutinin

  Publication DOI: 10.1016/0304-4165(88)90118-3
  Journal NLM ID: 0217513
  Publisher: Elsevier
  Institutions: Department of Chemistry, Osaka University College of Science, Osaka, Japan, Laboratory of Biochemistry, Faculty of Agriculture, Kyushu University, Fukuoka, Japan, Institute for Protein Research, Osaka University, Osaka Japan
  Methods: 1H NMR, HPLC, enzymatic digestion, partial acetolysis, RP-HPLC, CC
  
   
  
  Ricinus communis
  CSDB ID 109196 (all data & tools)
• Article ID: 10688
  Hase S, Koyama S, Daiyasu H, Takemoto H, Hara S, Kobayashi Y, Kyogoku Y, Ikenaka T "Structure of a sugar chain of a protease inhibitor isolated from Barbados pride (Caesalpinia pulcherrima Sw.) seeds" - Journal of Biochemistry 100 (1986) 1-10
  

  An asparagine-linked sugar chain of a protease inhibitor from barbados pride (Caesalpinia pulcherrima Sw.) was liberated by hydrazinolysis. After N-acetylation, the reducing end residue of this carbohydrate unit was coupled with 2-aminopyridine and the pyridylamino (PA-) derivative was purified by gel-filtration and reversed-phase HPLC. The structure of the resulting PA-sugar chain was determined mainly by stepwise exoglycosidase digestions and 500 MHz 1H-NMR spectroscopy and proved to be as follows: (formula; see text).

  NCBI PubMed ID: 3759923
  Journal NLM ID: 0376600
  Publisher: Japanese Biochemical Society
  
   
  
  Caesalpinia pulcherrima
  CSDB ID 132295 (all data & tools)

Show legend Show as text

Structure type: oligomer
Compound class: N-glycan
Contained glycoepitopes: IEDB_114701,IEDB_115005,IEDB_116644,IEDB_122244,IEDB_123886,IEDB_123887,IEDB_123888,IEDB_130701,IEDB_135813,IEDB_136045,IEDB_137340,IEDB_137485,IEDB_141793,IEDB_141807,IEDB_142489,IEDB_144562,IEDB_144983,IEDB_145668,IEDB_145669,IEDB_148491,IEDB_148492,IEDB_148493,IEDB_150092,IEDB_151531,IEDB_152206,IEDB_152214,IEDB_153212,IEDB_167188,IEDB_174332,IEDB_174333,IEDB_548907,IEDB_983930,SB_197,SB_198,SB_33,SB_44,SB_67,SB_72,SB_73,SB_74,SB_85,SB_86

• Article ID: 9546
  Takahashi N, Hotta T, Ishihara H, Mori M, Tejima S, Bligny R, Akazawa T, Endo S, Arata Y "Xylose-containing common structural unit in N-linked oligosaccharides of laccase from sycamore cells" - Biochemistry 25(2) (1986) 388-395
  

  The structures of asparagine-linked oligosaccharides of laccase excreted by sycamore (Acer pseudoplatanus L.) cells are reported. Peptic glycopeptides obtained from the laccase were treated with N-oligosaccharide glycopeptidase (EC 3.5.1.52) to release the oligosaccharide moieties. The oligosaccharides thus obtained were fractionated into six components by gel filtration, thin-layer chromatography, and high-performance liquid chromatography. The structures of the isolated oligosaccharides were determined by sugar analysis, exoglycosidase digestion, and methylation analysis in combination with high-resolution proton nuclear magnetic resonance spectroscopy. It was found that (1) the six oligosaccharides are a series of compounds of xylose-containing biantennary complex types that share as the core a common structural unit, i.e., Xyl-β(l-2)[Man-α(l-6)]Man-β(l-4)GlcNAc-β(l-4)[Fuc-α(1-3)]GlcNAc, and (2) mannose, N-acetylglucosamine, galactose, and fucose residues are additionally linked to the core as the outer chain moieties.

  Publication DOI: 10.1021/bi00350a018
  Journal NLM ID: 0370623
  Publisher: American Chemical Society
  Institutions: Department of Hygienic Chemistry, Faculty of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan, Department of Biochemistry, Nagoya City University Medical School, Nagoya, Japan, Centre National de la Recherche Scientifique, CENG, Grenoble, France, Research Institute for Biological Regulation School of Agriculture, Nagoya University, Nagoya, Japan, Department of Biophysics and Biochemistry, Faculty of Science, University of Tokyo, Tokyo, Japan
  Methods: 1H NMR, TLC, acid hydrolysis, HPLC, enzymatic digestion, methylation analysis
  
   
  
  Acer pseudoplatanus
  CSDB ID 107208 (all data & tools)

Show legend Show as text

Structure type: oligomer
Compound class: N-glycan
Contained glycoepitopes: IEDB_114701,IEDB_115005,IEDB_116644,IEDB_122244,IEDB_123886,IEDB_123887,IEDB_123888,IEDB_130701,IEDB_135813,IEDB_136045,IEDB_137340,IEDB_137485,IEDB_141793,IEDB_141807,IEDB_142489,IEDB_144562,IEDB_144983,IEDB_145668,IEDB_145669,IEDB_148491,IEDB_148492,IEDB_148493,IEDB_150092,IEDB_151531,IEDB_152206,IEDB_152214,IEDB_153212,IEDB_167188,IEDB_174332,IEDB_174333,IEDB_548907,IEDB_983930,SB_197,SB_198,SB_33,SB_44,SB_67,SB_72,SB_73,SB_74,SB_85,SB_86

• Article ID: 9546
  Takahashi N, Hotta T, Ishihara H, Mori M, Tejima S, Bligny R, Akazawa T, Endo S, Arata Y "Xylose-containing common structural unit in N-linked oligosaccharides of laccase from sycamore cells" - Biochemistry 25(2) (1986) 388-395
  

  The structures of asparagine-linked oligosaccharides of laccase excreted by sycamore (Acer pseudoplatanus L.) cells are reported. Peptic glycopeptides obtained from the laccase were treated with N-oligosaccharide glycopeptidase (EC 3.5.1.52) to release the oligosaccharide moieties. The oligosaccharides thus obtained were fractionated into six components by gel filtration, thin-layer chromatography, and high-performance liquid chromatography. The structures of the isolated oligosaccharides were determined by sugar analysis, exoglycosidase digestion, and methylation analysis in combination with high-resolution proton nuclear magnetic resonance spectroscopy. It was found that (1) the six oligosaccharides are a series of compounds of xylose-containing biantennary complex types that share as the core a common structural unit, i.e., Xyl-β(l-2)[Man-α(l-6)]Man-β(l-4)GlcNAc-β(l-4)[Fuc-α(1-3)]GlcNAc, and (2) mannose, N-acetylglucosamine, galactose, and fucose residues are additionally linked to the core as the outer chain moieties.

  Publication DOI: 10.1021/bi00350a018
  Journal NLM ID: 0370623
  Publisher: American Chemical Society
  Institutions: Department of Hygienic Chemistry, Faculty of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan, Department of Biochemistry, Nagoya City University Medical School, Nagoya, Japan, Centre National de la Recherche Scientifique, CENG, Grenoble, France, Research Institute for Biological Regulation School of Agriculture, Nagoya University, Nagoya, Japan, Department of Biophysics and Biochemistry, Faculty of Science, University of Tokyo, Tokyo, Japan
  Methods: 1H NMR, TLC, acid hydrolysis, HPLC, enzymatic digestion, methylation analysis
  
   
  
  Acer pseudoplatanus
  CSDB ID 107207 (all data & tools)

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Structure type: oligomer
Compound class: N-glycan
Contained glycoepitopes: IEDB_114701,IEDB_115005,IEDB_116644,IEDB_122244,IEDB_123886,IEDB_123887,IEDB_123888,IEDB_130701,IEDB_135813,IEDB_136045,IEDB_137340,IEDB_137485,IEDB_141793,IEDB_141807,IEDB_142489,IEDB_144562,IEDB_144983,IEDB_145668,IEDB_145669,IEDB_146665,IEDB_148491,IEDB_148492,IEDB_148493,IEDB_150092,IEDB_151531,IEDB_152206,IEDB_152214,IEDB_153212,IEDB_167186,IEDB_167188,IEDB_167189,IEDB_174332,IEDB_174333,IEDB_548907,IEDB_983930,SB_197,SB_198,SB_33,SB_44,SB_67,SB_72,SB_73,SB_74,SB_85,SB_86

• Article ID: 9546
  Takahashi N, Hotta T, Ishihara H, Mori M, Tejima S, Bligny R, Akazawa T, Endo S, Arata Y "Xylose-containing common structural unit in N-linked oligosaccharides of laccase from sycamore cells" - Biochemistry 25(2) (1986) 388-395
  

  The structures of asparagine-linked oligosaccharides of laccase excreted by sycamore (Acer pseudoplatanus L.) cells are reported. Peptic glycopeptides obtained from the laccase were treated with N-oligosaccharide glycopeptidase (EC 3.5.1.52) to release the oligosaccharide moieties. The oligosaccharides thus obtained were fractionated into six components by gel filtration, thin-layer chromatography, and high-performance liquid chromatography. The structures of the isolated oligosaccharides were determined by sugar analysis, exoglycosidase digestion, and methylation analysis in combination with high-resolution proton nuclear magnetic resonance spectroscopy. It was found that (1) the six oligosaccharides are a series of compounds of xylose-containing biantennary complex types that share as the core a common structural unit, i.e., Xyl-β(l-2)[Man-α(l-6)]Man-β(l-4)GlcNAc-β(l-4)[Fuc-α(1-3)]GlcNAc, and (2) mannose, N-acetylglucosamine, galactose, and fucose residues are additionally linked to the core as the outer chain moieties.

  Publication DOI: 10.1021/bi00350a018
  Journal NLM ID: 0370623
  Publisher: American Chemical Society
  Institutions: Department of Hygienic Chemistry, Faculty of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan, Department of Biochemistry, Nagoya City University Medical School, Nagoya, Japan, Centre National de la Recherche Scientifique, CENG, Grenoble, France, Research Institute for Biological Regulation School of Agriculture, Nagoya University, Nagoya, Japan, Department of Biophysics and Biochemistry, Faculty of Science, University of Tokyo, Tokyo, Japan
  Methods: 1H NMR, TLC, acid hydrolysis, HPLC, enzymatic digestion, methylation analysis
  
   
  
  Acer pseudoplatanus
  CSDB ID 107209 (all data & tools)