Information on EC 3.2.1.130 - glycoprotein endo-alpha-1,2-mannosidase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
3.2.1.130
-
RECOMMENDED NAME
GeneOntology No.
glycoprotein endo-alpha-1,2-mannosidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
hydrolysis of the terminal alpha-D-glucosyl-(1,3)-D-mannosyl unit from the GlcMan9(GlcNAc)2 oligosaccharide component of the glycoprotein produced in the Golgi membrane
show the reaction diagram
involved in the synthesis of glycoproteins
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-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of O-glycosyl bond
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-
-
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SYSTEMATIC NAME
IUBMB Comments
glycoprotein glucosylmannohydrolase
Involved in the synthesis of glycoproteins.
CAS REGISTRY NUMBER
COMMENTARY hide
108022-16-8
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125858-79-9
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
kangaroo rat
-
-
Manually annotated by BRENDA team
syrian or golden hamster
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
no activity in Cricetulus griseus
CD-strain
-
-
Manually annotated by BRENDA team
Wistar
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-
Manually annotated by BRENDA team
pig
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
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the alpha-endomannosidase gene is associated with panic disorder and social anxiety disorder
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(3R,4R,5R)-3-(alpha-D-mannopyranosyloxy)-4-hydroxy-5-(hydroxymethyl)-piperidine (alpha-Man-(1->3)-isofagomine) + H2O
?
show the reaction diagram
-
-
-
-
?
(3R,4R,5R)-3-(alpha-D-mannopyranosyloxy)-4-hydroxy-5-(hydroxymethyl)-piperidine + H2O
D-mannose + (3R,4R,5R)-3,4-dihydro-5-hydroxymethylpiperidine
show the reaction diagram
-
-
-
-
?
3-N-dansylaminopropyl [alpha-D-glucopyranosyl-(1->3)]-alpha-D-mannopyranosyl-(1->2)-alpha-D-mannopyranosyl (1->2)-alpha-D-mannopyranoside + H2O
3-N-dansylaminopropyl-alpha-D-mannopyranosyl-(1->2)-alpha-D-mannopyranoside + ?
show the reaction diagram
-
-
-
-
?
4-methylumbelliferyl (alpha-D-glucopyranosyl)-(1->3)-alpha-D-mannopyranoside + H2O
?
show the reaction diagram
4-methylumbelliferyl (alpha-D-mannopyranosyl)-(1->3)-alpha-D-mannopyranoside + H2O
?
show the reaction diagram
alpha-glucopyranosyl-(1->3)-alpha-mannopyranosyl fluoride + H2O
?
show the reaction diagram
alpha-mannan + H2O
?
show the reaction diagram
Glc(1->3)Man9GlcNAc2 + H2O
Man8GlcNAc + glucosyl alpha(1->3)-mannose
show the reaction diagram
Glc(1->3)Man9GlcNAc2 + H2O
Man8GlcNAc + glucosyl alpha1,3-mannose
show the reaction diagram
Glc1Man5GlcNAc1 + H2O
?
show the reaction diagram
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in general, endomannosidase variants preferentially process monoglucosylated oligosaccharides with truncated mannose chains, i.e. highest levels of activity are observed for Glc1Man5GlcNAc1 and Glc1Man7GlcNAc1/2
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-
?
Glc2Man9GlcNAc + H2O
?
show the reaction diagram
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at 14% the rate of GlcMan9GlcNAc hydrolysis
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-
?
Glc3Man9GlcNAc2 + H2O
?
show the reaction diagram
GlcMan5GlcNAc2 + H2O
alpha-D-glucosyl-1,3-D-mannopyranose + Man4GlcNAc2
show the reaction diagram
GlcMan7GlcNAc + H2O
?
show the reaction diagram
GlcMan7GlcNAc2 + H2O
Man6GlcNAc2 + glucosyl alpha(1->3)-D-mannose
show the reaction diagram
GlcMan7GlcNAc2 + H2O
Man6GlcNAc2 + glucosyl alpha-(1->3)-D-mannose
show the reaction diagram
GlcMan8GlcNAc + H2O
?
show the reaction diagram
GlcMan9GlcNAc + H2O
Man8GlcNAc + glucosyl alpha1,3-mannose
show the reaction diagram
GlcMan9GlcNAc2 + H2O
?
show the reaction diagram
human alpha1-antitrypsin + H2O
?
show the reaction diagram
methyl alpha-D-mannopyranosyl-(1->3)-alpha-D-mannopyranosyl-(1->2)-alpha-D-mannopyranosyl-(1->2)-alpha-D-mannopyranoside + H2O
alpha-(1->3)-D-mannobiose + methyl alpha-(1->2)-D-mannobioside + ?
show the reaction diagram
vesicular stomatitis virus G protein + H2O
?
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
GlcMan7GlcNAc2 + H2O
Man6GlcNAc2 + glucosyl alpha(1->3)-D-mannose
show the reaction diagram
GlcMan7GlcNAc2 + H2O
Man6GlcNAc2 + glucosyl alpha-(1->3)-D-mannose
show the reaction diagram
GlcMan9GlcNAc + H2O
Man8GlcNAc + glucosyl alpha1,3-mannose
show the reaction diagram
GlcMan9GlcNAc2 + H2O
?
show the reaction diagram
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(3R,4R,5R)-3-(alpha-D-glucopyranosyloxy)-4-hydroxy-5-(hydroxymethyl)piperidine
alpha-glucopyranosyl-(1->3)-deoxymannojirimycin
alpha-mannopyranosyl-(1->3)-isofagomine
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-
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alpha-mannopyranosyl-1->3-isofagomine
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-
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Glucosyl alpha1-3(1-deoxy)mannojirimycin
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-
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Triton X-100
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requirement, membrane-bound enzyme
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.404
4-methylumbelliferyl (alpha-D-glucopyranosyl)-(1->3)-alpha-D-mannopyranoside
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wild type enzyme, in 25 mM HEPES pH 7.0, 100 mM NaCl, at 37C
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0.038 - 1.827
4-methylumbelliferyl (alpha-D-mannopyranosyl)-(1->3)-alpha-D-mannopyranoside
0.48 - 2.6
alpha-glucopyranosyl-(1->3)-alpha-mannopyranosyl fluoride
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0000717
4-methylumbelliferyl (alpha-D-glucopyranosyl)-(1->3)-alpha-D-mannopyranoside
Bacteroides xylanisolvens
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wild type enzyme, in 25 mM HEPES pH 7.0, 100 mM NaCl, at 37C
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0.00008 - 0.00115
4-methylumbelliferyl (alpha-D-mannopyranosyl)-(1->3)-alpha-D-mannopyranoside
0.58 - 6.34
alpha-glucopyranosyl-(1->3)-alpha-mannopyranosyl fluoride
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.78 - 102.7
4-methylumbelliferyl (alpha-D-glucopyranosyl)-(1->3)-alpha-D-mannopyranoside
0.208 - 6.3
4-methylumbelliferyl (alpha-D-mannopyranosyl)-(1->3)-alpha-D-mannopyranoside
0.2 - 9.9
alpha-glucopyranosyl-(1->3)-alpha-mannopyranosyl fluoride
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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specific activity 10.855 units/mg, 1 unit is the amount of enzyme that catalyzes the release of 1000 dpm of glucosyl-alpha-3Man in 1 h
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.2 - 6.8
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-
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 9
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about half-maximal activity at pH 6 and 8.5
5.5 - 7.5
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about half-maximal activity at pH 5.8 and 7.7
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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baby hamster kidney cell line served as host for vesicular stomatitis virus
Manually annotated by BRENDA team
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thymoma cell line served as host for vesicular stomatitis virus
Manually annotated by BRENDA team
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no activity due to molecular defect in endomannosidase, CHO cells have a W188C substitution, Trp188 is critical for Golgi localization and in vivo activity of endomannosidase
Manually annotated by BRENDA team
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surface epithelium and crypt epithelium
Manually annotated by BRENDA team
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villus epithelium and crypt epithelium
Manually annotated by BRENDA team
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cell line served as host for vesicular stomatitis virus
Manually annotated by BRENDA team
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kidney cell line served as host for vesicular stomatitis virus
Manually annotated by BRENDA team
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cell line serves as host for vesicular stomatitis virus
Manually annotated by BRENDA team
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cell line serves as host for vesicular stomatitis virus
Manually annotated by BRENDA team
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seminiferous tubules and Leydig cells
Manually annotated by BRENDA team
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follicular cells
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
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its cytoplasmic domain is required for its efficient Golgi localization. Proper topology rather than the presence of positively charged amino acids in the cytoplasmic tail is critical for Golgi localization of rat endomannosidase
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
52000
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cDNA analysis
53000
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ER-retained CHO cell endomannosidase, immunoprecipitation/Western blot analysis
53220
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sequence analysis
56000
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SDS-PAGE
58000
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Golgi-localized rat endomannosidase species, immunoprecipitation/Western blot analysis
61000
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Golgi-localized rat endomannosidase species, immunoprecipitation/Western blot analysis
380000
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gel filtration
560000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oligomer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphoprotein
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phosphorylation is occurring in the Golgi apparatus
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
single wavelength anomalous dispersion technique
wild type and mutant enzyme E333Q, in complex with 4-methylumbelliferyl (alpha-D-glucopyranosyl)-(1->3)-alpha-D-mannopyranoside or alpha-mannopyranosyl-(1->3)-isofagomine, hanging drop vapor diffusion method, using 2.8 M sodium acetate pH 6.4
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-70C, isolated membranes, storage for 40 days with several freeze-thawings results in about a 30% loss in activity
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-80C, in intact Golgi membranes, 10% loss of activity within 6 months
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-80C, purified enzyme, at 0.03 mg protein/ml pH 6.8, 50% loss of activity within 6 months, increased stability at 0.5 mg protein/ml, bovine serum albumin stabilizes
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
NiSO4-charged HiTrap column chromatography and S75 gel filtration
recombinant protein, expressed in Escherichia coli
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cDNA expressed in Escherichia coli
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cloning of endomannosidase from CHO-K1 cells, amplicon subcloned into the Hind III/Xho I sites of the expression vector pcDNA6/myc-His, in frame with the C-terminal myc- and His-tag. Myc-tagged CHO endomannosidase expressed in CHO Lec23 cells. Generation of rat/CHO endomannosidase hybrid constructs
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endomannosidase subcloned into the HindIII/XhoI site of the pcDNA6 vector in frame with the myc-tag or into the Hind III/Bam HI site of the pEGFP vector for endomannosidase, expressed in CHO-K1 cells. CHO-K1 cells expressing DELTAsig-rEndo or DELTATMD-rEndo
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expressed in CHO-K1 cells
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expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli JM109
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expression in Pichia pastoris
myc-tagged rat endomannosidase expressed in CHO Lec23 cells, generation of rat/CHO endomannosidase hybrid constructs
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subcloned into the expression vector pcDNA6/myc-His, overexpression in clone 9 cells and castanospermine-treated hepatocytes
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the level of alpha-endomannosidase mRNA is reduced to 89%, 54% and 34% at 2, 5 and 10 h, respectively, after the addition of actinomycin D
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E154A
the mutant shows near zero activity
E329A
the mutant shows near zero activity
E332A
the mutant shows approximate 50fold decrease in catalytic activity with respect to wild type enzyme
E154A
-
the mutant shows near zero activity
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E329A
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the mutant shows near zero activity
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E332A
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the mutant shows approximate 50fold decrease in catalytic activity with respect to wild type enzyme
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C188A
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enzymatic activity can not be fully rescued with a back-engineered Cys188Ala CHO cell endomannosidase
C188S
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enzymatic activity can not be fully rescued with a back-engineered Cys188Ser CHO cell endomannosidase
C188W
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enzyme activity can be reversed in cells with a back-engineered Cys188Trp CHO cell endomannosidase, in particular N-glycans of alpha1-antitrypsin become fully processed
additional information
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in CHO-K1 cells expressing DELTAsig-rEndo, lacking the whole signal sequence, or DELTATMD-rEndo, lacking the transmembrane domain, endomannosidase is undetectable. DELTACT-rEndo, lacking the cytoplasmic tail, exhibits an ER localization and fails to maintain a type II membrane topology
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
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cocaine-induced paranoia is associated with 6 single-nucleotide polymorphisms in European American families and 9 single-nucleotide polymorphisms in African American families. Association of MANEA single-nucleotide polymorphisms with cocaine dependence in both family samples is much weaker. The A allele of the 3' untranslated region single-nucleotide polymorphism rs9387522 is associated with increased risk of cocaine-induced paranoia