Information on EC 2.4.1.50 - procollagen galactosyltransferase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.4.1.50
-
RECOMMENDED NAME
GeneOntology No.
procollagen galactosyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
UDP-alpha-D-galactose + procollagen 5-hydroxy-L-lysine = UDP + procollagen 5-(D-galactosyloxy)-L-lysine
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexosyl group transfer
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Lysine degradation
-
-
Other types of O-glycan biosynthesis
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-
SYSTEMATIC NAME
IUBMB Comments
UDP-galactose:procollagen-5-hydroxy-L-lysine D-galactosyltransferase
Probably involved in the synthesis of carbohydrate units in complement (cf. EC 2.4.1.66 procollagen glucosyltransferase).
CAS REGISTRY NUMBER
COMMENTARY hide
9028-07-3
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
denatured collagen type I + UDP-alpha-D-galactose
UDP + galactosylated collagen type I
show the reaction diagram
-
-
-
-
?
UDP-alpha-D-galactose + procollagen 5-hydroxy-L-lysine
UDP + procollagen 5-(D-galactosyloxy)-L-lysine
show the reaction diagram
UDP-galactose + bovine collagen type I (deglycosylated)
UDP + ?
show the reaction diagram
-
-
-
?
UDPgalactose + alpha1-glycoprotein with sialic acid and galactose removed
?
show the reaction diagram
-
less active than with guinea pig skin collagen with sugar residues removed
-
-
?
UDPgalactose + fetuin with sialic acid and galactose removed
?
show the reaction diagram
-
33% of activity with bovine achilles tendon collagen with glucose and galactose residues removed
-
-
?
UDPgalactose + ichthyocol
?
show the reaction diagram
-
fish collagen ichthyocol as best substrate, peptides derived from collagenase- and pronase-digestion of ichthyocol are less effective
-
-
?
UDPgalactose + ovalbumin
?
show the reaction diagram
-
17.9% of activity with bovine achilles tendon collagen with glucose and galactose residues removed
-
-
?
UDPgalactose + procollagen 5-hydroxy-L-lysine
UDP + procollagen 5-(D-galactosyloxy)-L-lysine
show the reaction diagram
UDPgalactose + submaxillary glycoprotein with sialic acid removed
?
show the reaction diagram
additional information
?
-
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
UDP-alpha-D-galactose + procollagen 5-hydroxy-L-lysine
UDP + procollagen 5-(D-galactosyloxy)-L-lysine
show the reaction diagram
UDPgalactose + procollagen 5-hydroxy-L-lysine
UDP + procollagen 5-(D-galactosyloxy)-L-lysine
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cd2+
-
20% of activation by Mg2+
Fe2+
-
activates, 2.7% as effective as Mn2+
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Co2+
-
6 mM: 50% inhibition
concanavalin A
-
Cu2+
-
-
Galactosylhydroxylysine
-
-
gelatinized calf skin collagen
-
above 60 mg/ml
-
human renal basement membrane collagen
-
glucose-free
-
hydroxylysine
-
weak
Mn2+
-
above 2.5 mM
p-hydroxymercuribenzoate
-
10 mM, 50% inhibition
p-mercuribenzoate
-
strong, 0.007 mM: 50% inhibition
Pb2+
-
-
UDPgalacturonate
-
-
UDPglucose
-
-
UDPglucuronate
-
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
dithiothreitol
Nonidet P-40
-
activates
Triton X-100
Urea
-
2 mM, activates
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.028
calf skin collagen
-
value based on actual amount of galactosylhydroxylysyl-residues in calf skin collagen
-
0.1 - 0.2
hydroxylysyl acceptor sites
-
-
-
0.28
ichthyocol
-
value based on actual amount of galactosylhydroxylysyl-residues in ichthyocol
-
0.01877 - 0.03353
UDP-galactose
0.02 - 0.049
UDPgalactose
additional information
additional information
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00093
-
-
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 6.5
-
2 pH-optima: 6-6.5 and 7.5-8
7 - 7.4
-
-
7
-
about
7.5 - 8
-
2 pH-optima: 6-6.5 and 7.5-8
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 9.5
-
about half-maximal activity at pH 5.5 and 9.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
embryo bone
Manually annotated by BRENDA team
-
fetal lung diploid fibroblasts, cell lines WI-38 and IMR-90
Manually annotated by BRENDA team
-
fetal lung diploid fibroblasts, WI-38 and IMR-90 cells
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
presumed location within
-
Manually annotated by BRENDA team
-
associated with membrane structures
Manually annotated by BRENDA team
-
blood platelet plasma membrane, may be buried in the interior of the membrane or located on its inner surface
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50000
-
3 species of enzyme activity: 450000, 200000 and 50000 Da, gel filtration
80000
-
value about, Western blot
85000
-
SDS-PAGE
200000
-
3 species of enzyme activity: 450000, 200000 and 50000 Da, gel filtration
450000
-
3 species of enzyme activity: 450000, 200000 and 50000 Da, possibly an aggregate of enzyme or enzyme with other proteins, gel filtration
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
gel filtration inactivates
-
highly labile enzyme, DTT stabilizes during purification, 0.25 M or 0.1 M galactose or glycerol 25% v/v do not stabilize
-
unstable during purification
-
unstable in crude extracts
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ethylene glycol
-
50% v/v, stable to
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 50% v/v ethylene glycol, several weeks, stable
-
4°C, unstable to storage, urea, galactose, Triton X-100, glycerol or ethylene glycol do not stabilize
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
1000fold; partial
-
250-300fold; optimal extraction requires high concentrations of detergent and salt
-
50-150fold; partial
-
; Ni-NTA-agarose
-
affinity chromatography
-
partial
-
partial, solubilized with Triton X-100
-
skin, partial
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
chimeric proteins (GLT25D1-cerebral endothelial cell adhesion molecule (CEECAM1)) expressed in SF-9 cell
-
epitope-tagged proteins expressed in human hepatoma cell line Huh7
-
expressed in MEF cells
-
overexpression in HT-1080 cells, expression of fragments in Escherichia coli; wild-type protein and His-tagged version expressed in fibrosarcoma cells HT-1080, 30 kDa His-tagged amino-terminal fragment (amino acid residue 25-290) with glycosyltransferase activity expressed in Escherichia coli
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recombinant baculoviruses are produced in Spodoptera frugiperda Sf9 cells; recombinant baculoviruses are produced in Spodoptera frugiperda Sf9 cells
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
PLOD2 mRNA is induced greater than 4fold by hypoxia within 12 h and remains elevated for 72 hours of continuous hypoxia
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A187D/A189D
-
30 kDa His-tagged amino-terminal fragment with mutations shows 2% glycosyltransferase activity of wild type fragment
D166A/D168A
-
inactive protein
D207H
-
the mutation is associated with skin fragility, delayed wound healing, joint hyperlaxity and contractures, muscle hypotonia, intellectual disability, and a spondyloepimetaphyseal dysplasia with bone fragility and severe kyphoscoliosis
D336S
-
lightly decreased activity
D461A/D463A
-
inactive protein
D585A/D587A
-
wild type activity
G217S
-
the mutation is associated with skin fragility, delayed wound healing, joint hyperlaxity and contractures, muscle hypotonia, intellectual disability, and a spondyloepimetaphyseal dysplasia with bone fragility and severe kyphoscoliosis
P292N
-
lightly decreased activity
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
high enzyme expression in breast cancer biopsies is associated with increased risk of mortality