Information on EC 2.4.2.38 - glycoprotein 2-beta-D-xylosyltransferase

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The expected taxonomic range for this enzyme is: Eukaryota

EC NUMBER
COMMENTARY
2.4.2.38
-
RECOMMENDED NAME
GeneOntology No.
glycoprotein 2-beta-D-xylosyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
UDP-D-xylose + N4-{N-acetyl-beta-D-glucosaminyl-(1,2)-alpha-D-mannosyl-(1,3)-[N-acetyl-beta-D-glucosaminyl-(1,2)-alpha-D-mannosyl-(1,6)]-beta-D-mannosyl-(1,4)-N-acetyl-beta-D-glucosaminyl-(1,4)-N-acetyl-beta-D-glucosaminyl}asparagine = UDP + N4-{N-acetyl-beta-D-glucosaminyl-(1,2)-alpha-D-mannosyl-(1,3)-[N-acetyl-beta-D-glucosaminyl-(1,2)-alpha-D-mannosyl-(1,6)]-[beta-D-xylosyl-(1,2)]-beta-D-mannosyl-(1,4)-N-acetyl-beta-D-glucosaminyl-(1,4)-N-acetyl-beta-D-glucosaminyl}asparagine
show the reaction diagram
-
-
-
-
UDP-D-xylose + N4-{N-acetyl-beta-D-glucosaminyl-(1-2)-alpha-D-mannosyl-(1-3)-[N-acetyl-beta-D-glucosaminyl-(1-2)-alpha-D-mannosyl-(1-6)]-beta-D-mannosyl-(1-4)-N-acetyl-beta-D-glucosaminyl-(1-4)-N-acetyl-beta-D-glucosaminyl}asparagine = UDP + N4-{N-acetyl-beta-D-glucosaminyl-(1-2)-alpha-D-mannosyl-(1-3)-[N-acetyl-beta-D-glucosaminyl-(1-2)-alpha-D-mannosyl-(1-6)]-[beta-D-xylosyl-(1-2)]-beta-D-mannosyl-(1-4)-N-acetyl-beta-D-glucosaminyl-(1-4)-N-acetyl-beta-D-glucosaminyl}asparagine
show the reaction diagram
the catalytic domain of XylT encompasses residues Lys57Cys534
-
UDP-D-xylose + N4-{N-acetyl-beta-D-glucosaminyl-(1->2)-alpha-D-mannosyl-(1->3)-[N-acetyl-beta-D-glucosaminyl-(1->2)-alpha-D-mannosyl-(1->6)]-beta-D-mannosyl-(1->4)-N-acetyl-beta-D-glucosaminyl-(1->4)-N-acetyl-beta-D-glucosaminyl}asparagine = UDP + N4-{N-acetyl-beta-D-glucosaminyl-(1->2)-alpha-D-mannosyl-(1->3)-[N-acetyl-beta-D-glucosaminyl-(1->2)-alpha-D-mannosyl-(1->6)]-[beta-D-xylosyl-(1->2)]-beta-D-mannosyl-(1->4)-N-acetyl-beta-D-glucosaminyl-(1->4)-N-acetyl-beta-D-glucosaminyl}asparagine
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
transfer of glycosyl group
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Various types of N-glycan biosynthesis
-
-
SYSTEMATIC NAME
IUBMB Comments
UDP-D-xylose:glycoprotein (D-xylose to the 3,6-disubstituted mannose of N4-{N-acetyl-beta-D-glucosaminyl-(1->2)-alpha-D-mannosyl-(1->3)-[N-acetyl-beta-D-glucosaminyl-(1->2)-alpha-D-mannosyl-(1->6)]-beta-D-mannosyl-(1->4)-N-acetyl-beta-D-glucosaminyl-(1->4
Specific for N-linked oligosaccharides (N-glycans).
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
beta-1,2-xylosyltransferase
-
-
-
-
beta-1,4-mannosylglycoprotein beta-1,2-xylosyltransferase
-
-
-
-
XylT
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
141256-56-6
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
serotype A (H99); serotype D
-
-
Manually annotated by BRENDA team
serotype D, strain JEC21 (serotype D MATalpha) and strain ags1delta (serotype D MATalpha ags1delta)
UniProt
Manually annotated by BRENDA team
serotype D, wild-type strain JEC21 (serotype D MATalpha) and mutant strain JEC21/cxt1delta (serotype D, isogenic MAT alpha, encapsulated and nourseothricin-resistant, lacking CXT1p)
UniProt
Manually annotated by BRENDA team
strains JEC20, JEC21 and CAP67
UniProt
Manually annotated by BRENDA team
wild-type strain JEC20 (serotype D MAT a), wild-type strain JEC21 (serotype D MAT alpha), mutant strain CAP67 (serotype D MAT alpha cap59), mutant strain JEC21/cxt1delta (serotype D, isogenic MAT alpha, encapsulated and nourseothricin-resistant, lacking CXT1p), mutant strain CAP67/cxt1delta (serotype D, isogenic MAT alpha, acapsulated and nourseothricin-resistant, lacking CXT1p)
-
-
Manually annotated by BRENDA team
isozyme Xylt1
SwissProt
Manually annotated by BRENDA team
no activity in Spodoptera frugiperda
-
-
-
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
gene xt1; cv. Monalisa
SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
Q5K8R6
loss of XT1 does not affect in vitro growth of mutant cells or general morphology of their capsules. The two main capsule polysaccharides, glucuronoxylomannan and galactoxylomannan, both are missing beta1,2-xylose residues. Deletion of XT1 leads to attenuation of cryptococcal growth in a mouse model of infection
malfunction
-
expression of the tomato beta1,2-xylosyltransferase is down-regulated by RNA interference in transgenic tomato plants. On the basis of IgE immunoblotting assays XylT-RNAi tomato fruits reveal a patient-specific reduction in IgE reactivity. Patients with tomato allergy have reduced mean wheal diameters in skin prick tests, demonstrating a reduced allergenic potential of XylT-RNAi tomato fruits in vivo
physiological function
Q5K8R6
Xt1p has a defined role in capsule biosynthesis
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(Xylbeta1-2GnGn)-GP + UDP-xylose
?
show the reaction diagram
-
i.e. GnGnX
-
-
?
pyridylaminated-GnGn glycopeptide + UDP-xylose
?
show the reaction diagram
Q2UVB3
-
-
-
?
UDP-D-xylose + alpha-D-mannosyl-(1,6)-[N-acetyl-beta-D-glucosaminyl-(1,2)-alpha-D-mannosyl-(1,3)]-beta-D-mannosyl-O-(CH2)7-CH3
UDP + ?
show the reaction diagram
-
-
-
-
?
UDP-D-xylose + D-mannosyl-alpha-(1,3)-D-mannose
UDP + D-xylosyl-beta-(1,2)-D-mannosyl-alpha-(1,3)-D-mannose
show the reaction diagram
Q5K8R6
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, 20C
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
-
?
UDP-D-xylose + D-mannosyl-alpha-(1,3)-D-mannose
UDP + D-xylosyl-beta-(1,2)-D-mannosyl-alpha-(1,3)-D-mannose
show the reaction diagram
Q5K8R6
membranes, radiolabelled UDP-xylose, alpha-(1,3)-mannobiose (resembles backbone or dimannose moiety of natural substrates)
product: trisaccharide with xylose moiety linked to reducing mannose of alpha-(1,3)-mannobiose, radiolabel detection by scintillation counting or thin layer chromatography and autoradiography, product purification for NMR and glycosyl linkage analyses
-
?
UDP-D-xylose + galactoxylomannan
UDP + [beta-D-xylose-(1,2)]-galactoxylomannan
show the reaction diagram
Q5K8R6
-
JEC21/cxt1delta and CAP67/cxt1delta lacking CXT1p: almost complete loss of beta-xylose residues (1,2)-linked to mannose, unchanged amounts of beta-xylose (1,3)-linked to galactose
-
?
UDP-D-xylose + galactoxylomannan
UDP + [beta-D-xylosyl-(1,2)]-galactoxylomannan
show the reaction diagram
Q5K8R6
-
-
-
?
UDP-D-xylose + GlcNAcbeta1-2Manalpha1-3Manbeta1-O-(CH2)7-CH3
UDP + GlcNAcbeta1-2Manalpha1-3(Xylbeta1-2)Manbeta1-O-(CH2)7-CH3
show the reaction diagram
-
-
-
?
UDP-D-xylose + GlcNAcbeta1-2Manalpha1-6(Galbeta1-4GlcNAcbeta1-2Manalpha1-3)Manbeta1-4GlcNAc
UDP + GlcNAcbeta1-2Manalpha1-6(Galbeta1-4GlcNAcbeta1-2Manalpha1-3)(Xylbeta1-2)Manbeta1-4GlcNAc
show the reaction diagram
-
-
-
?
UDP-D-xylose + GlcNAcbeta1-2Manalpha1-6(Galbeta1-4GlcNAcbeta1-2Manalpha1-3)Manbeta1-O-(CH2)7-CH3
UDP + GlcNAcbeta1-2Manalpha1-6(Galbeta1-4GlcNAcbeta1-2Manalpha1-3)(Xylbeta1-2)Manbeta1-O-(CH2)7-CH3
show the reaction diagram
-
-
-
?
UDP-D-xylose + GlcNAcbeta1-2Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)Manbeta1-4GlcNAcbeta1-4(Fucalpha1-6)GlcNAc1-O-(N-benzyloxycarbonyl)-Tyr
UDP + GlcNAcbeta1-2Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)(Xylbeta1-2)Manbeta1-4GlcNAcbeta1-4(Fucalpha1-6)GlcNAc1-O-(N-benzyloxycarbonyl)-Tyr
show the reaction diagram
-
-
-
?
UDP-D-xylose + GlcNAcbeta1-2Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)Manbeta1-4GlcNAcbeta1-4GlcNAc
UDP + GlcNAcbeta1-2Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)(Xylbeta1-2)Manbeta1-4GlcNAcbeta1-4GlcNAc
show the reaction diagram
-
-
-
?
UDP-D-xylose + GlcNAcbeta1-2Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)Manbeta1-4GlcNAcbeta1-4GlcNAc-Asn
UDP + GlcNAcbeta1-2Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)(Xylbeta1-2)Manbeta1-4GlcNAcbeta1-4GlcNAc-Asn
show the reaction diagram
-
-
-
?
UDP-D-xylose + GlcNAcbeta1-2Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)Manbeta1-4GlcNAcbeta1-4GlcNAc1-O-(N-benzyloxycarbonyl)-Tyr
UDP + GlcNAcbeta1-2Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)(Xylbeta1-2)Manbeta1-4GlcNAcbeta1-4GlcNAc1-O-(N-benzyloxycarbonyl)-Tyr
show the reaction diagram
-
-
-
?
UDP-D-xylose + GlcNAcbeta1-2Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)Manbeta1-4GlcNAcbeta1-4GlcNAc1-O-(pyrid-2-yl)amine
UDP + GlcNAcbeta1-2Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)(Xylbeta1-2)Manbeta1-4GlcNAcbeta1-4GlcNAc1-O-(pyrid-2-yl)amine
show the reaction diagram
-
-
-
?
UDP-D-xylose + GlcNAcbeta1-2Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)Manbeta1-4GlcNAcbeta1-4GlcNAc1-O-(pyrid-2-yl)amine
UDP + GlcNAcbeta1-2Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)(Xylbeta1-2)Manbeta1-4GlcNAcbeta1-4GlcNAc1-O-(pyrid-2-yl)amine
show the reaction diagram
Q9LDH0
-
-
?
UDP-D-xylose + GlcNAcbeta1-2Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)Manbeta1-O-(CH2)7-CH3
UDP + GlcNAcbeta1-2Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)(Xylbeta1-2)Manbeta1-O-(CH2)7-CH3
show the reaction diagram
-
-
-
?
UDP-D-xylose + glucuronoxylomannan
UDP + [beta-D-xylose-(1,2)]-glucuronoxylomannan
show the reaction diagram
Q5K8R6
-
JEC21/cxt1delta and CAP67/cxt1delta lacking CXT1p: 30% decrease in amount of xylose, 39% decrease in O-2 xylose-substituted mannose, 31.5% increase in O-2-unsubstituted mannose, loss of 1/3 of beta-(1,2)-linked xylose, revealed by NMR analyses
-
?
UDP-D-xylose + glucuronoxylomannan
UDP + [beta-D-xylosyl-(1,2)]-glucuronoxylomannan
show the reaction diagram
Q5K8R6
-
-
-
?
UDP-D-xylose + Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)Manbeta1-4GlcNAcbeta1-4GlcNAc1-O-(N-benzyloxycarbonyl)-Tyr
UDP + Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)(Xylbeta1-2)Manbeta1-4GlcNAcbeta1-4GlcNAc1-O-(N-benzyloxycarbonyl)-Tyr
show the reaction diagram
-
-
-
?
UDP-D-xylose + Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)Manbeta1-O-(CH2)7-CH3
UDP + Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)(Xylbeta1-2)Manbeta1-O-(CH2)7-CH3
show the reaction diagram
-
-
-
?
UDP-D-xylose + N4-[N-acetyl-beta-D-glucosaminyl-(1,2)-alpha-D-mannosyl-(1,3)-[N-acetyl-beta-D-glucosaminyl-(1,2)-alpha-D-mannosyl-(1,6)]-beta-D-mannosyl-(1,4)-N-acetyl-beta-D-glucosaminyl-(1,4)-N-acetyl-beta-D-glucosaminyl]asparagine
UDP + N4-[N-acetyl-beta-D-glucosaminyl-(1,2)-alpha-D-mannosyl-(1,3)-[N-acetyl-beta-D-glucosaminyl-(1,2)-alpha-D-mannosyl-(1,6)]-[beta-D-xylosyl-(1,2)]-beta-D-mannosyl-(1,4)-N-acetyl-beta-D-glucosaminyl-(1,4)-N-acetyl-beta-D-glucosaminyl]asparagine
show the reaction diagram
-
-
-
-
?
UDP-xylose + dansylated Galbeta-(1,4)-GlcNAcbeta-(1,2)-Manalpha-(1,6)-(Galbeta-(1,4)-GlcNAcbeta-(1,2)-Manalpha-(1,3))-Manbeta-(1,4)-GlcNAcbeta-(1,4)-GlcNAc
UDP + ?
show the reaction diagram
-
i.e. dansylated GnGn-peptide
-
-
?
UDP-xylose + dansylated Galbeta-(1,4)-GlcNAcbeta-(1,2)-Manalpha-(1,6)-(Galbeta-(1,4)-GlcNAcbeta-(1,2)-Manalpha-(1,3))-Manbeta-(1,4)-GlcNAcbeta-(1,4)-GlcNAc
UDP + ?
show the reaction diagram
Q6ZFH6
i.e. dansylated GnGn-peptide
-
-
?
UDP-xylose + mannosyl-alpha-(1,3)-mannosyl-alpha-(1,4)-galactosyl-beta-(1,6)-mannosyl-alpha-(1,2)-inositol phosphorylceramide
UDP + mannosyl-alpha-(1,3)-[xylosyl-beta-(1,2)]-mannosyl-alpha-(1,4)-galactosyl-beta-(1,6)-mannosyl-alpha-(1,2)-inositol phosphorylceramide
show the reaction diagram
Q5K8R6
no transfer of xylose residues onto substrate in absence of Cxt1p (JEC21/cxt1delta) as revealed by mass spectrometry and HPTLC, mannosyl-alpha-(1,3)-mannose-alpha motif also present in substrates for capsule polysaccharide synthesis
core structure of glycosylinositol phosphorylceramides, GIPCs
-
?
UDP-xylose + mannosyl-alpha-(1,6)-mannosyl-alpha-(1,3)-mannosyl-alpha-(1,4)-galactosyl-beta-(1,6)-mannosyl-alpha-(1,2)-inositol phosphorylceramide
UDP + mannosyl-alpha-(1,6)-mannosyl-alpha-(1,3)-[xylosyl-beta-(1,2)]-mannosyl-alpha-(1,4)-galactosyl-beta-(1,6)-mannosyl-alpha-(1,2)-inositol phosphorylceramide
show the reaction diagram
Q5K8R6
no transfer of xylose residues onto substrate in absence of Cxt1p (JEC21/cxt1delta) as revealed by mass spectrometry and HPTLC
-
-
?
Xylbeta1-2[Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)]Manbeta1-O-octyl + UDP-xylose
?
show the reaction diagram
-
i.e. MGnX-octyl
-
-
?
Xylbeta1-2[Manalpha1-6(Manalpha1-3)]Manbeta1-O-octyl + UDP-xylose
?
show the reaction diagram
-
i.e. MMX-octyl
-
-
?
[GlcNAcbeta1-2Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)Manbeta1-4GlcNAcbeta1-4(Fucalpha1-3)GlcNAc]-GP + UDP-xylose
?
show the reaction diagram
-
i.e. GnGnF
-
-
?
[Manalpha1-6(Manalpha1-3)Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)Manbeta1-4GlcNAcbeta1-4GlcNAc]-GP + UDP-xylose
?
show the reaction diagram
-
i.e. Man5Gn
-
-
?
GlcNAcbeta1-2Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)Manbeta1-O-octyl + UDP-xylose
?
show the reaction diagram
-
i.e. GnGn-octyl
-
-
?
additional information
?
-
-
-
strongly reduced biosynthesis of beta 1,2-xylose-containing glycoepitopes upon knock-down of endogenous beta 1,2-xylosyltransferase using RNA interference approaches
-
-
additional information
?
-
-
-
strongly reduced biosynthesis of beta 1,2-xylose-containing glycoepitopes upon knock-down of endogenous beta 1,2-xylosyltransferase using sense, antisense and RNA interference approaches
-
-
additional information
?
-
-
-
human anti HIV monoclonal antibody 2G12 produced in absence of endogenous beta 1,2-xylosyltransferase (knock-down by RNA interference) is functional but contains N-glycan species lacking beta 1,2-xylose residues corresponding to authentic human N-glycosylation
-
-
additional information
?
-
-
probably involved in the biosynthesis of the inner part of hemocyanin glycans
-
-
-
additional information
?
-
Q6ZFH6
enzyme is involved in N-glycan biosynthesis, quantification and structure analysis of N-glycans in rice leaves
-
-
-
additional information
?
-
-
substrate specificity, substrates need to possess a beta(1,2)-linked GlcNAc residue at the Man-alpha-(1,3) arm, Man-alpha-(1,6)-(Man-alpha(1,3))-Man-beta(1-R) is no acceptor substrate, integrity of a large domain located between amino acid 31 and the C-terminal lumenal region is required for activity, the cytosolic tail and the transmembrane region are not essential for activity, but the stem region and the C-terminal end are
-
-
-
additional information
?
-
-
transfer of D-xylose from UDP-D-xylose to the core beta-linked mannose of N-linked oligosaccharides
-
-
-
additional information
?
-
-
the enzyme is involved in the biosynthesis of glycoprotein-bound N-glycans in plants and acts at multiple stages of the plant N-glycosylation pathway, overview
-
-
-
additional information
?
-
Q2UVB3
the enzyme is involved in the modification of asparagine-linked oligosaccharides of maize glycoproteins with beta1,2-xylose sugar residues
-
-
-
additional information
?
-
-
the enzyme is specific for UDP-xylose, no activity with Man5-GP and MM-octyl, acceptor substrate specificity, overview
-
-
-
additional information
?
-
Q2UVB3
the recombinant enzyme is able to convert a mammalian acceptor glycoprotein into an antigen binding anti-plant N-glycan antibodies, overview
-
-
-
additional information
?
-
Q5K8R6
preferred substrate depends on wild-type strain, strain JEC21: both products detectable (mannosyl-alpha-(1,6)-mannosyl-alpha-(1,3)-[xylosyl-beta-(1,2)]-mannosyl-alpha-(1,4)-galactosyl-beta-(1,6)-mannosyl-alpha-(1,2)-inositol phosphorylceramide and mannosyl-alpha-(1,3)-[xylosyl-beta-(1,2)]-mannosyl-alpha-(1,4)-galactosyl-beta-(1,6)-mannosyl-alpha-(1,2)-inositol phosphorylceramide) at a ratio of approximately 1:1
-
-
-
additional information
?
-
Q5K8R6
transfer of beta (1,2)-xylose, capsule synthesis
-
-
-
additional information
?
-
Q5K8R6
transfer of beta (1,2)-xylose, synthesis of capsule polysaccharides and fungi-specific glycosphingolipids
-
-
-
additional information
?
-
Q5K8R6
lack of CXT1p in membranes from JEC21/cxt1delta and CAP67/cxt1delta leads to dramatic reduction in D-xylose-beta-(1,2)-D-mannose-alpha-(1,3)-D-mannose synthesising activity in presence of exogenously added substrate
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
UDP-D-xylose + galactoxylomannan
UDP + [beta-D-xylose-(1,2)]-galactoxylomannan
show the reaction diagram
Q5K8R6
-
JEC21/cxt1delta and CAP67/cxt1delta lacking CXT1p: almost complete loss of beta-xylose residues (1,2)-linked to mannose, unchanged amounts of beta-xylose (1,3)-linked to galactose
-
?
UDP-D-xylose + galactoxylomannan
UDP + [beta-D-xylosyl-(1,2)]-galactoxylomannan
show the reaction diagram
Q5K8R6
-
-
-
?
UDP-D-xylose + glucuronoxylomannan
UDP + [beta-D-xylose-(1,2)]-glucuronoxylomannan
show the reaction diagram
Q5K8R6
-
JEC21/cxt1delta and CAP67/cxt1delta lacking CXT1p: 30% decrease in amount of xylose, 39% decrease in O-2 xylose-substituted mannose, 31.5% increase in O-2-unsubstituted mannose, loss of 1/3 of beta-(1,2)-linked xylose, revealed by NMR analyses
-
?
UDP-D-xylose + glucuronoxylomannan
UDP + [beta-D-xylosyl-(1,2)]-glucuronoxylomannan
show the reaction diagram
Q5K8R6
-
-
-
?
UDP-D-xylose + N4-[N-acetyl-beta-D-glucosaminyl-(1,2)-alpha-D-mannosyl-(1,3)-[N-acetyl-beta-D-glucosaminyl-(1,2)-alpha-D-mannosyl-(1,6)]-beta-D-mannosyl-(1,4)-N-acetyl-beta-D-glucosaminyl-(1,4)-N-acetyl-beta-D-glucosaminyl]asparagine
UDP + N4-[N-acetyl-beta-D-glucosaminyl-(1,2)-alpha-D-mannosyl-(1,3)-[N-acetyl-beta-D-glucosaminyl-(1,2)-alpha-D-mannosyl-(1,6)]-[beta-D-xylosyl-(1,2)]-beta-D-mannosyl-(1,4)-N-acetyl-beta-D-glucosaminyl-(1,4)-N-acetyl-beta-D-glucosaminyl]asparagine
show the reaction diagram
-
-
-
-
?
UDP-xylose + mannosyl-alpha-(1,3)-mannosyl-alpha-(1,4)-galactosyl-beta-(1,6)-mannosyl-alpha-(1,2)-inositol phosphorylceramide
UDP + mannosyl-alpha-(1,3)-[xylosyl-beta-(1,2)]-mannosyl-alpha-(1,4)-galactosyl-beta-(1,6)-mannosyl-alpha-(1,2)-inositol phosphorylceramide
show the reaction diagram
Q5K8R6
no transfer of xylose residues onto substrate in absence of Cxt1p (JEC21/cxt1delta) as revealed by mass spectrometry and HPTLC, mannosyl-alpha-(1,3)-mannose-alpha motif also present in substrates for capsule polysaccharide synthesis
core structure of glycosylinositol phosphorylceramides, GIPCs
-
?
UDP-xylose + mannosyl-alpha-(1,6)-mannosyl-alpha-(1,3)-mannosyl-alpha-(1,4)-galactosyl-beta-(1,6)-mannosyl-alpha-(1,2)-inositol phosphorylceramide
UDP + mannosyl-alpha-(1,6)-mannosyl-alpha-(1,3)-[xylosyl-beta-(1,2)]-mannosyl-alpha-(1,4)-galactosyl-beta-(1,6)-mannosyl-alpha-(1,2)-inositol phosphorylceramide
show the reaction diagram
Q5K8R6
no transfer of xylose residues onto substrate in absence of Cxt1p (JEC21/cxt1delta) as revealed by mass spectrometry and HPTLC
-
-
?
additional information
?
-
-
-
strongly reduced biosynthesis of beta 1,2-xylose-containing glycoepitopes upon knock-down of endogenous beta 1,2-xylosyltransferase using RNA interference approaches
-
-
additional information
?
-
-
-
strongly reduced biosynthesis of beta 1,2-xylose-containing glycoepitopes upon knock-down of endogenous beta 1,2-xylosyltransferase using sense, antisense and RNA interference approaches
-
-
additional information
?
-
-
probably involved in the biosynthesis of the inner part of hemocyanin glycans
-
-
-
additional information
?
-
Q6ZFH6
enzyme is involved in N-glycan biosynthesis, quantification and structure analysis of N-glycans in rice leaves
-
-
-
additional information
?
-
-
the enzyme is involved in the biosynthesis of glycoprotein-bound N-glycans in plants and acts at multiple stages of the plant N-glycosylation pathway, overview
-
-
-
additional information
?
-
Q2UVB3
the enzyme is involved in the modification of asparagine-linked oligosaccharides of maize glycoproteins with beta1,2-xylose sugar residues
-
-
-
additional information
?
-
Q5K8R6
preferred substrate depends on wild-type strain, strain JEC21: both products detectable (mannosyl-alpha-(1,6)-mannosyl-alpha-(1,3)-[xylosyl-beta-(1,2)]-mannosyl-alpha-(1,4)-galactosyl-beta-(1,6)-mannosyl-alpha-(1,2)-inositol phosphorylceramide and mannosyl-alpha-(1,3)-[xylosyl-beta-(1,2)]-mannosyl-alpha-(1,4)-galactosyl-beta-(1,6)-mannosyl-alpha-(1,2)-inositol phosphorylceramide) at a ratio of approximately 1:1
-
-
-
additional information
?
-
Q5K8R6
transfer of beta (1,2)-xylose, capsule synthesis
-
-
-
additional information
?
-
Q5K8R6
transfer of beta (1,2)-xylose, synthesis of capsule polysaccharides and fungi-specific glycosphingolipids
-
-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Ca2+
Q6ZFH6
activates, enzyme requires divalent cations for full activity
Fe2+
Q6ZFH6
activates, enzyme requires divalent cations for full activity
Mg2+
Q6ZFH6
activates, enzyme requires divalent cations for full activity
Mn2+
-
6.25-12.5 mM, stimulation
Mn2+
-
highly required for activity
Mn2+
Q6ZFH6
activates, enzyme requires divalent cations for full activity
additional information
-
no requirements for metal ions
additional information
Q2UVB3
is active in the presence of 10 mM EDTA and does not require the addition of any metal ions for activity
additional information
Q5K8R6
activity not dependent on divalent cations
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Co2+
Q6ZFH6
-
Co2+
-
59% inhibition at 10 mM
Cu2+
Q6ZFH6
-
EDTA
-
strong inhibition
EDTA
Q6ZFH6
-
EDTA
-
20% inhibition at 10 mM
Ni2+
-
99% inhibition at 10 mM
UDP
-
competitive
UDP-glucose
-
-
Zn2+
Q6ZFH6
-
Zn2+
-
95% inhibition at 10 mM
Mn2+
Q2UVB3
above 10 mM
additional information
-
substrate inhibition
-
additional information
Q5K8R6
10 mM EDTA does not reduce endogenous activity
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.055
dansylated Galbeta-(1,4)-GlcNAcbeta-(1,2)-Manalpha-(1,6)-(Galbeta-(1,4)-GlcNAcbeta-(1,2)-Manalpha-(1,3))-Manbeta-(1,4)-GlcNAcbeta-(1,4)-GlcNAc
Q6ZFH6
recombinant soluble enzyme, pH 6.0, 16C
0.13
UDP-xylose
-
pH 7.0, 37C, recombinant mutant DELTA39 XylT
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.042
UDP
-
pH 7.0, 37C, recombinant mutant DELTA39 XylT
4.9
UDP-glucose
-
pH 7.0, 37C, recombinant mutant DELTA39 XylT
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.000007
-
-
0.33
-
purified recombinant enzyme
additional information
-
activity of truncated enzyme mutants
additional information
Q5K8R6
transfer of 300 cpm/min/mg by endogenous activity in crude membranes; transfer of 890000 cpm/min/mg by endogenous, purified activity after alpha-(1,3)-mannobiose affinity chromatography
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.5 - 7
-
recombinant mutant DELTA39 XylT
6.5
-
assay at
6.7
Q6ZFH6
-
7
Q2UVB3
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.5 - 8.5
-
negligible activity at pH 5.0
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
16
Q6ZFH6
-
37
Q2UVB3
assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
Q6ZFH6
low activity at 30C
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
Q2UVB3
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
enzyme possesses a transmembrane domain
-
Manually annotated by BRENDA team
-
the CTS region, formed by the N-terminal 90 amino acids, is responsible for correct targeting of the enzyme to the cytosplamic membrane
Manually annotated by BRENDA team
-
enzyme possesses a transmembrane domain
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
55000 - 59000
-
gel filtration, SDS-PAGE
394593
62000
-
calculation from sequence
394592
79000
Q5K8R6
calculated, 694 amino acids; hexa-His tagged mutant N141Q, 12% SDS-PAGE
692993
90000
Q5K8R6
hexa-His tagged wild-type and mutants D550A and D659A, 12% SDS-PAGE; purified endogenous activity (serotype D), 12% SDS-PAGE and S-300 gel filtration, identity confirmed by mass spectrometry
692993
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
Q2UVB3
x * 56000, recombinant enzyme, SDS-PAGE
?
-
x * 60000, recombinant enzyme, SDS-PAGE
additional information
-
primary sequence and hydrophobicity analysis of recombinant XylT
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
glycoprotein
-
3 potential N-glycosylation sites, Asn51, Asn301, and Asn478, 2 of which are glycosylated, i.e. Asn51 and Asn301, occupancy of at least one site is necessary for enzyme activity and stability, glycosylation sites of diverse mutants, overview
glycoprotein
-
three potential N-glycosylation sites of the wild-type enzyme as well as of the recombinant enzyme are occupied by N-linked oligosaccharides, N-glycosylation is not essential for proper folding of XylT
proteolytic modification
-
proteolytic processing of recombinant XylT occurs between amino acids Arg178 and Arg179, a cleavage sequence compatible with the enzymatic properties of the baculoviral cysteine proteinase
side-chain modification
Q5K8R6
N-linked glycosylation site
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.5 - 7.5
-
sensitive to changes in pH of storage solution
394593
additional information
-
the purified enzyme is stable for prolonged periods at neutral pH and at temperatures of up to 30C
671831
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30
-
the purified enzyme is stable for prolonged periods at neutral pH and at temperatures of up to 30C
671831
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
the purified recombinant enzyme is not stable to a single freeze-thaw cycle
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4C, purified recombinant enzyme, 3 weeks stable without loss of activity
-
4C to -20 C, HEPES-buffer, pH 7.0, 10% glycerol, 0.2% Triton X-100, 1 mM dithiothreitol, 90 days, 20%
-
4C, purified recombinant enzyme, 3 weeks stable without loss of activity
Q6ZFH6
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant N-terminally His5-tagged enzyme from Pichia pastoris cell medium by nickel chelating affinity chromatography and ultrafiltration to homogeneity
-
subcellular fractionation
-
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, 4C), 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
Q5K8R6
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, expression of full length wild-type enzyme and diverse mutants in Spodoptera frugiperda Sf9 insect cells via baculovirus infection system as soluble proteins lacking the transmembrane domain, expression of the full length enzyme and enzyme fragments as GFP-fusion proteins in tobacco cells Golgi medial cisternae via Agrobacterium tumefaciens infection system for study of enzyme subcellular localization
-
expression in insect cells
-
expression of N-terminally His5-tagged enzyme in Pichia pastoris, recombinant enzyme is secreted to the medium
-
expression of the enzyme's isolated CTS region, N-terminal 90 amino acids, and of the T and C region separately, as fusion protein with GFP in Nicotiana benthamiana leaf epidermal cells, the CT region, but not only the T or C regions, is sufficient to sustain Golgi retention without the involvement of any luminal sequences
-
expression of wild-type and mutant enzymes in Spodoptera frugiperda SF21 cells using the baculovirus infection system
-
recombinant AtXYLT is expressed in insect cells
-
5-kb PCR product cloned into the TOPO pCR2.1 vector and maintained in Escherichia coli DH5alpha; amplification of the gene with flanking regions (1.3 kb upstream of the start codon, 1 kb downstream of the gene) from serotype D (strain JEC21) genomic DNA for cloning in pCR2.1TOPO, generation of replacement cassettes for replacing endogenous CXT1p: (i) nourseothricin acetyltransferase resistance gene flanked by the CXT1p flanking regions, transformed as linear DNA in CAP67 strain, (ii) cxt1 gene lacking 3 two-thirds of the encoding sequence flanked by its flanking regions, transformed as linear DNA in JEC21 strain
Q5K8R6
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
Q5K8R6
gene xylt, transfection of chimeric constructs via Agrobacterium tumefaciens strain C58Z707, overview
Q0PMC9
DNA and amino acid sequence determination and analysis, gene is located on chromosome 8, functional expression of the enzyme in Pichia pastoris as soluble protein, secretion to the cell culture medium
Q6ZFH6
DNA and amino acid sequence determination and analysis, genetic organization and structure, functional expression in Spodoptera frugiperda Sf21 cells using the baculovirus transfection system
Q2UVB3
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
S53A
-
site-directed mutagenesis, possesses the N-glycosylation sites Asn301 and Asn478
S53A/T303A
-
site-directed mutagenesis
T303A
-
site-directed mutagenesis, possesses the N-glycosylation sites Asn51 and Asn478
T480A
-
site-directed mutagenesis, possesses the N-glycosylation sites Asn51 and Asn301
D550A
Q5K8R6
mutation of first DXD motif to AXD, complete loss of activity, expression in Saccharomyces cerevisiae
D659A
Q5K8R6
mutation of second DXD motif to AXD, dramatic reduction in activity, expression in Saccharomyces cerevisiae
additional information
-
construction of transgenic Nicotiana benthamiana plants expressing the CTSregion, T and C region, and a membrane-shed enzyme version, the latter is secreted to the apoplast, the T or C region constructs are located in the cytoplasm, the CTS construct is located in the cytoplasmic membrane, overview
additional information
-
serial deletions at both the N- and the C-terminal ends, integrity of a large domain located between amino acid 31 and the C-terminal lumenal region is required for activity, construction of transgenic tobacco expressing the enzyme via Agrobacterium tumefaciens infection system
additional information
-
construction of several N-terminal deletion mutants, the first 54 residues can be deleted without affecting the catalytic activity of the enzyme, removal of an additional five amino acids lead to the formation of an inactive protein
N141Q
Q5K8R6
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
additional information
Q0PMC9
co-expression of human monoclonal antibodies with specific RNAi to block beta-1,2-xylosyltransferase and alpha-1,3-fucosyltransferase prevents unwanted plant specific N-glycosylation of the recombinant antibody proteins, method optimization, overview
additional information
Q2UVB3
targeted downregulation of the enzyme and the alpha1,3-fucosyltransferase allow production of heterologous proteins in a maize expression system
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
Q5K8R6
development of diagnostic and therapeutic strategies due to importance of xylose transfer in cryptococcal biology, pathogenesis and virulence
medicine
Q5K8R6
xylose residues within the capsule polysaccharide are important for normal host-pathogen interaction, loss of CXT1p attenuates virulence
medicine
Q5K8R6
loss of XT1 leads to attenuation of virulence
biotechnology
Q0PMC9
the plant is useful for expression of human monoclonal antibodies free of zoonotic pathogens, co-expression of RNAi to block beta-1,2-xylosyltransferase and alpha-1,3-fucosyltransferase prevents unwanted plant specific N-glycosylation of the recombinant antibody proteins, overview
biotechnology
-
pharmaceutical protein production in plant, glycoengineering: humanized N-glycosylation in plants by knock-down of plant-specific N-glycan maturation enzymes
biotechnology
-
glycoengineering: humanized N-glycosylation in plants by knock-down of plant-specific N-glycan maturation enzymes, pharmaceutical glycoprotein production in plant
synthesis
Q2UVB3
potential of maize and the beta1,2-xylosyltransferase as a production system for heterologous glycoproteins