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Literature summary extracted from

  • Klutts, J.S.; Levery, S.B.; Doering, T.L.
    A beta-1,2-xylosyltransferase from Cryptococcus neoformans defines a new family of glycosyltransferases (2007), J. Biol. Chem., 282, 17890-17899.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.2.38 from JEC21 cDNA by PCR with primers introducing C-terminal hexa-His tag, in pCR2.1 TOPO for sequencing, in pPGK (cloned between promoter and terminator of phosphoglycerate kinase) for expression in Saccharomyces cerevisiae that lacks endogenous xylosyltransferase activity Cryptococcus neoformans

Protein Variants

EC Number Protein Variants Comment Organism
2.4.2.38 D550A mutation of first DXD motif to AXD, complete loss of activity, expression in Saccharomyces cerevisiae Cryptococcus neoformans
2.4.2.38 D659A mutation of second DXD motif to AXD, dramatic reduction in activity, expression in Saccharomyces cerevisiae Cryptococcus neoformans
2.4.2.38 N141Q mutation of the N-glycosylation site, molecular weight shift to 79 kDa corresponding to the molecular weight of the unmodified enzyme, no change in activity, expression in Saccharomyces cerevisiae Cryptococcus neoformans

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.4.2.38 additional information 10 mM EDTA does not reduce endogenous activity Cryptococcus neoformans

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.4.2.38 additional information activity not dependent on divalent cations Cryptococcus neoformans

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.2.38 79000
-
 calculated, 694 amino acids Cryptococcus neoformans
2.4.2.38 79000
-
 hexa-His tagged mutant N141Q, 12% SDS-PAGE Cryptococcus neoformans
2.4.2.38 90000
-
 hexa-His tagged wild-type and mutants D550A and D659A, 12% SDS-PAGE Cryptococcus neoformans
2.4.2.38 90000
-
 purified endogenous activity (serotype D), 12% SDS-PAGE and S-300 gel filtration, identity confirmed by mass spectrometry Cryptococcus neoformans

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.2.38 additional information Cryptococcus neoformans transfer of beta (1,2)-xylose, capsule synthesis ?
-
 ?
2.4.2.38 UDP-D-xylose + galactoxylomannan Cryptococcus neoformans
-
 UDP + [beta-D-xylosyl-(1,2)]-galactoxylomannan
-
 ?
2.4.2.38 UDP-D-xylose + glucuronoxylomannan Cryptococcus neoformans
-
 UDP + [beta-D-xylosyl-(1,2)]-glucuronoxylomannan
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.4.2.38 Aspergillus clavatus
-
-
-
2.4.2.38 Aspergillus fischeri
-
-
-
2.4.2.38 Aspergillus fumigatus
-
-
-
2.4.2.38 Aspergillus nidulans
-
-
-
2.4.2.38 Aspergillus terreus
-
-
-
2.4.2.38 Chaetomium globosum
-
-
-
2.4.2.38 Coccidioides immitis
-
-
-
2.4.2.38 Cryptococcus neoformans
-
 serotype A (H99)
-
2.4.2.38 Cryptococcus neoformans
-
 serotype D
-
2.4.2.38 Cryptococcus neoformans Q5K8R6 serotype D, strain JEC21 (serotype D MATalpha) and strain ags1delta (serotype D MATalpha ags1delta)
-
2.4.2.38 Fusarium graminearum
-
-
-
2.4.2.38 Neurospora crassa
-
-
-
2.4.2.38 Pyricularia grisea
-
-
-
2.4.2.38 Ustilago maydis
-
-
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
2.4.2.38 side-chain modification N-linked glycosylation site Cryptococcus neoformans

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.2.38 endogenous activity (2967-fold purification) from serotype D ags1delta strain: solubilisation from crude membranes with 1% Triton X-100 (on ice, 30 min), HiPrep 16/10 Q-Sepharose chromatography with NaCl gradient elution (pH 8), activity peaks were pooled and concentrated (spin, 10000 Da cut-off), HiPrep 26/60 Sephacryl S-300 gel filtration chromatography with isocratic gradient elution (pH 8), concanavalin A-Sepharose 4B chromatography with alpha-methyl-mannoside elution (incubation over night, 4°C), HiTrap DEAE chromatorgraphy with NaCl gradient elution (pH 8), alpha-(1,3)-mannobiose agarose resin affinity chromatography with NaCl gradient (pH 6.5), activity peaks were combined and concentrated, overexpressed hexa-His tagged enzyme: solubilisation, Talon Superflow cobalt metal affinity chromatography, elution with 250 mM imidazole, pH 7.5 Cryptococcus neoformans

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.4.2.38 additional information
-
 transfer of 300 cpm/min/mg by endogenous activity in crude membranes Cryptococcus neoformans
2.4.2.38 additional information
-
 transfer of 890000 cpm/min/mg by endogenous, purified activity after alpha-(1,3)-mannobiose affinity chromatography Cryptococcus neoformans

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.2.38 additional information transfer of beta (1,2)-xylose, capsule synthesis Cryptococcus neoformans ?
-
 ?
2.4.2.38 UDP-D-xylose + D-mannosyl-alpha-(1,3)-D-mannose membranes or purified enzyme, radiolabelled UDP-xylose, 8.5-12.5 mM alpha-(1,3)-mannobiose (resembles backbone or dimannose moiety of natural substrates), pH 6.5, 4 h, 20°C Cryptococcus neoformans UDP + D-xylosyl-beta-(1,2)-D-mannosyl-alpha-(1,3)-D-mannose product: trisaccharide with xylose moiety linked to reducing mannose of alpha-(1,3)-mannobiose as revealed by mass spectrometry, NMR and methylation linkage analyses, radiolabel detection by scintillation counting or thin layer chromatography and autoradiography or product purification by solid phase extraction ?
2.4.2.38 UDP-D-xylose + galactoxylomannan
-
 Cryptococcus neoformans UDP + [beta-D-xylosyl-(1,2)]-galactoxylomannan
-
 ?
2.4.2.38 UDP-D-xylose + glucuronoxylomannan
-
 Cryptococcus neoformans UDP + [beta-D-xylosyl-(1,2)]-glucuronoxylomannan
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.4.2.38 CAP3 previous name based on homology to CAP10 Cryptococcus neoformans
2.4.2.38 cryptococcus xylosyltransferase 1
-
 Cryptococcus neoformans
2.4.2.38 CXT1
-
 Cryptococcus neoformans
2.4.2.38 Cxt1p
-
 Cryptococcus neoformans
2.4.2.38 CXT1p homolog 12 to 23% identity to CTX1p from Cryptococcus neoformans serotype D Aspergillus nidulans
2.4.2.38 CXT1p homolog 12 to 23% identity to CTX1p from Cryptococcus neoformans serotype D Neurospora crassa
2.4.2.38 CXT1p homolog 12 to 23% identity to CTX1p from Cryptococcus neoformans serotype D Aspergillus fischeri
2.4.2.38 CXT1p homolog 12 to 23% identity to CTX1p from Cryptococcus neoformans serotype D Aspergillus terreus
2.4.2.38 CXT1p homolog 12 to 23% identity to CTX1p from Cryptococcus neoformans serotype D Ustilago maydis
2.4.2.38 CXT1p homolog 12 to 23% identity to CTX1p from Cryptococcus neoformans serotype D Pyricularia grisea
2.4.2.38 CXT1p homolog 12 to 23% identity to CTX1p from Cryptococcus neoformans serotype D Aspergillus clavatus
2.4.2.38 CXT1p homolog 12 to 23% identity to CTX1p from Cryptococcus neoformans serotype D Aspergillus fumigatus
2.4.2.38 CXT1p homolog 12 to 23% identity to CTX1p from Cryptococcus neoformans serotype D Fusarium graminearum
2.4.2.38 CXT1p homolog 12 to 23% identity to CTX1p from Cryptococcus neoformans serotype D Chaetomium globosum
2.4.2.38 CXT1p homolog 12 to 23% identity to CTX1p from Cryptococcus neoformans serotype D Coccidioides immitis
2.4.2.38 CXT1p homolog group of 5 genes (CAP1, CAP5, CAP2, CAP4, CAP10) with 20 to 30% identity to CTX1p Cryptococcus neoformans