Information on EC 2.4.1.131 - GDP-Man:Man3GlcNAc2-PP-dolichol alpha-1,2-mannosyltransferase

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The expected taxonomic range for this enzyme is: Eukaryota

EC NUMBER
COMMENTARY hide
2.4.1.131
-
RECOMMENDED NAME
GeneOntology No.
GDP-Man:Man3GlcNAc2-PP-dolichol alpha-1,2-mannosyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
2 GDP-alpha-D-mannose + D-Man-alpha-(1->3)-[D-Man-alpha-(1->6)]-D-Man-beta-(1->4)-D-GlcNAc-beta-(1->4)-D-GlcNAc-diphosphodolichol = 2 GDP + D-Man-alpha-(1->2)-D-Man-alpha-(1->2)-D-Man-alpha-(1->3)-[D-Man-alpha-(1->6)]-D-Man-beta-(1->4)-D-GlcNAc-beta-(1->4)-D-GlcNAc-diphosphodolichol
show the reaction diagram
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-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexosyl group transfer
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
dolichyl-diphosphooligosaccharide biosynthesis
-
-
Metabolic pathways
-
-
N-Glycan biosynthesis
-
-
protein N-glycosylation (eukaryotic, high mannose)
-
-
Various types of N-glycan biosynthesis
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-
SYSTEMATIC NAME
IUBMB Comments
GDP-alpha-D-mannose:D-Man-alpha-(1->3)-[D-Man-alpha-(1->6)]-D-Man-beta-(1->4)-D-GlcNAc-beta-(1->4)-D-GlcNAc-diphosphodolichol 2-alpha-D-mannosyltransferase
The biosynthesis of asparagine-linked glycoproteins (N-linked protein glycosylation) utilizes a dolichyl diphosphate-linked glycosyl donor, which is assembled by the series of membrane-bound glycosyltransferases that comprise the dolichol pathway. ALG11 mannosyltransferase from Saccharomyces cerevisiae carries out two sequential steps in the formation of the lipid-linked core oligosaccharide, adding two mannose residues in alpha(1->2) linkages to the nascent oligosaccharide.
CAS REGISTRY NUMBER
COMMENTARY hide
74506-43-7
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
cow
-
-
Manually annotated by BRENDA team
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SwissProt
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2 GDP-D-mannose + D-Man-alpha-(1->3)-(D-Man-alpha-(1->6))-D-Man-beta-(1->4)-D-GlcNAc-beta-(1->4)-D-GlcNAc-diphosphodolichol
D-Man-alpha-(1->2)-D-Man-alpha-(1->2)-D-Man-alpha-(1->3)-(D-Man-alpha-(1->6))-D-Man-beta-(1->4)-D-GlcNAc-beta-(1->4)-D-GlcNAc-diphosphodolichol
show the reaction diagram
-
-
-
-
?
2 GDP-D-mannose + D-Man-alpha-(1->3)-[D-Man-alpha-(1->6)]-D-Man-beta-(1->4)-D-GlcNAc-beta-(1->4)-D-GlcNAc-diphosphodolichol
2 GDP + D-Man-alpha-(1->2)-D-Man-alpha-(1->2)-D-Man-alpha-(1->3)-[D-Man-alpha-(1->6)]-D-Man-beta-(1->4)-D-GlcNAc-beta-(1->4)-D-GlcNAc-diphosphodolichol
show the reaction diagram
GDP-mannose + lipid-linked Manalpha(1-3)Manbeta-GlcNAcbeta-GlcNAc
GDP + lipid-linked Manalpha(1-2)Manalpha(1-2)Manalpha(1-3)(Manalpha(1-6))Manbeta-GlcNAcbeta-GlcNAc
show the reaction diagram
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reaction via penta- and hexasaccharide intermediates to a lipid-linked heptasaccharide, presumably multiple mannosyltransferases involved
-
?
GDP-mannose + oligosaccharide-lipid acceptor
GDP + alpha-mannosyl-1,2-oligosaccharide-lipid
show the reaction diagram
GDPmannose + oligosaccharide-lipid acceptor
GDP + alpha-mannosyl-1,2-oligosaccharide-lipid
show the reaction diagram
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
2 GDP-D-mannose + D-Man-alpha-(1->3)-[D-Man-alpha-(1->6)]-D-Man-beta-(1->4)-D-GlcNAc-beta-(1->4)-D-GlcNAc-diphosphodolichol
2 GDP + D-Man-alpha-(1->2)-D-Man-alpha-(1->2)-D-Man-alpha-(1->3)-[D-Man-alpha-(1->6)]-D-Man-beta-(1->4)-D-GlcNAc-beta-(1->4)-D-GlcNAc-diphosphodolichol
show the reaction diagram
GDPmannose + oligosaccharide-lipid acceptor
GDP + alpha-mannosyl-1,2-oligosaccharide-lipid
show the reaction diagram
additional information
?
-
-
involved in asparagine-linked glycosylation, reaction proceeds in the endoplasmatic reticulum, role of the ALG9 step in downstream ER and Golgi N-glycan processing events
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ADP
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2 mM, less than 15% inhibition
ATP
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2 mM, less than 15% inhibition
Ca2+
-
weak
Cu2+
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strong
deoxycholate
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-
GDP
-
0.02 mM, 84% inhibition; 2 mM, 84% inhibition
GMP
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0.02 mM, 30% inhibition; 2 mM, 30% inhibition
GTP
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0.02 mM, 40% inhibition; 2 mM, 40% inhibition
Mn2+
-
-
taurocholate
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UDP
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2 mM, less than 15% inhibition
UDP-galactose
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2 mM, less than 15% inhibition
UDP-glucose
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2 mM, less than 15% inhibition
UDP-N-acetylglucosamine
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2 mM, less than 15% inhibition
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
EDTA
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slight stimulation
Endogen lipids
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soluble in CHCl3/CH3OH, 2:1, activation
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00064
D-Man-alpha-(1->3)-(D-Man-alpha-(1->6))-D-Man-beta-(1->4)-D-GlcNAc-beta-(1->4)-D-GlcNAc-diphosphodolichol
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pH 7.0, 37°C
0.0047
GDP-D-mannose
0.00064
oligosaccharide-lipid acceptor
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based on the assumption of incorporation of only a single mannosyl unit into the product
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.8 - 7.6
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broad
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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intimal layer
Manually annotated by BRENDA team
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expressed especially in developing stems and flowers
Manually annotated by BRENDA team
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lactating mammary tissue
Manually annotated by BRENDA team
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expressed especially in developing stems and flowers
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
63140
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calculated from sequence
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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x * 63143, calculated from sequence
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
no glycoprotein
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the protein is predicted to contain four N-linked glycosylation sites, but none appear to be utilized, because Alg11p is insensitive to digestion with endo H
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
0-4°C, solubilized enzyme, at least 2 weeks, stable
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0°C, partially purified enzyme, 5 days, stable
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4°C, solubilized enzyme, 2 weeks: about 40% loss of activity, 7 days: about 20% loss of activity
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
partial; solubilized with 0.15% v/v Nonidet P-40, partial
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ALG9 gene encodes a middle-arm terminal alpha1,2-mannosyltransferase Alg9p
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construction of a plasmid pREP1+/-ALG11HA for the expression of Saccharomyces cerevisiae ALG11 in Schizosaccharomyces pombe and introducing it into gmd3 mutant. The gmd3 mutant, carrying pREP+/-ALG11HA, grows at 37°C, whereas the gmd3 mutant, carrying vector pREP1 alone, does not. The defect in acid phosphatase glycosylation of gmd3 mutant is also suppressed by Saccharomyces cerevisiae ALG11. The gmd3+/alg11+ gene is a functional homologue of the Saccharomyces cerevisiae ALG11 gene
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overexpression in Escherichia coli. Two Alg2 constructs are expressed and isolated, one with the N-terminal TRX domain and C-terminal His and V5 epitope tags and the other with only an N-terminal His tag
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retroviral expression of human wild-type and mutated ALG11 cDNA in patient-derived fibroblasts as well as using a yeast alg11 deletion strain as a heterologous expression system for hALG11 variants
transformation of the LEW3 gene into the yeast mutant alg11, which exhibits a lowtemperature-sensitive phenotype and grows slowly at and below 30°C but normally at 37°C. Expression of LEW3 partially complements the slow-growth phenotype of the yeast mutant alg11, suggesting that the plant ALG11 homolog is able to partially substitute for the function of ALG11 in yeast
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
L85A
no effect on Alg11 mannosyltransferase activity
L86P
mutant enzyme does not complement the growth deffect of the mutant strain DELTAalgg3 (Saccharomyces cerevisiae)
L86S
mutant enzyme slightly complements the growth deffect of the mutant strain DELTAalgg3 (Saccharomyces cerevisiae)
E318A
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mutation has no effect on activity
E356A
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mutation has no effect on activity
E405/H406A
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double mutant no longer supports growth, the loss of activity is not caused by a failure of Alg11 expression
E405A/E413
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double mutant no longer supports growth, the loss of activity is not caused by a failure of Alg11 expression
E405A/E413A
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completely inactive mutant enzyme
F407A
-
mutation is not detrimental for growth, impairment of glycosylation of the vacuolar model glycoproteincarboxypeptidase Y
G84A
-
mutation has no effect on growth and glycosylation of the vacuolar model glycoproteincarboxypeptidase Y
G84P
-
mutation reduces growth and glycosylation of the vacuolar model glycoproteincarboxypeptidase Y
G85A
-
mutation has no effect on growth and glycosylation of the vacuolar model glycoproteincarboxypeptidase Y
G85P
-
mutation reduces growth and glycosylation of the vacuolar model glycoproteincarboxypeptidase Y
G87A
-
mutation has no effect on growth and glycosylation of the vacuolar model glycoproteincarboxypeptidase Y
G87P
-
mutation reduces growth and glycosylation of the vacuolar model glycoproteincarboxypeptidase Y
H406A
-
mutation is not detrimental for growth, impairment of glycosylation of the vacuolar model glycoproteincarboxypeptidase Y
K302A
-
mutation has no effect on activity
K319A
-
mutation causes loss of Alg11 activity
K322A
-
mutation has no effect on activity
K343A
-
mutation has no effect on activity
V412A
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mutation is not detrimental for growth