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Information on EC 3.2.1.46 - galactosylceramidase and Organism(s) Mus musculus and UniProt Accession P54818
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EC Tree
3.2.1.46 galactosylceramidaseIUBMB Comments
cf. EC 3.2.1.62 glycosylceramidase.
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Word Map
- 3.2.1.46
- krabbe
- leukodystrophy
- lysosomal
-
globoid
-
twitcher
- psychosine
- demyelination
- myelin
-
infantile
- galactosylsphingosine
- nerve
- neurodegenerative
- oligodendrocyte
-
hematopoietic
- sphingolipids
-
spastic
- arylsulfatase
-
saposins
-
metachromatic
- medicine
-
presymptomatic
-
schwann
- glycosphingolipids
- sphingomyelinase
- galactolipids
- gaucher
- gangliosidosis
- prosaposin
-
later-onset
- drug development
-
myelin-forming
-
late-infantile
- gm1-gangliosidosis
- glucosylceramidase
- glucocerebrosidase
-
gm1-ganglioside
- diagnostics
-
4-methylumbelliferyl
The taxonomic range for the selected organisms is: Mus musculus
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
Synonyms
galactocerebrosidase, galactosylceramidase, galactosylceramide beta-galactosidase, beta-galactocerebrosidase, galactocerebroside beta-galactosidase, gm1 ganglioside beta-galactosidase, lactosylceramidase i, cerebroside beta-galactosidase, beta-galactosylceramidase, galactosylceramidase i, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
beta-galactocerebrosidase
beta-galactosylceramidase
-
-
-
-
ceramidase, galacatosyl-
-
-
-
-
ceramide galactosidase
-
-
-
-
cerebroside beta-galactosidase
-
-
-
-
cerebroside galactosidase
-
-
-
-
galactoceramidase
-
-
-
-
galactocerebrosidase
galactocerebroside beta-galactosidase
-
-
-
-
galactocerebroside galactosidase
-
-
-
-
galactocerebroside-beta-D-galactosidase
-
-
-
-
Galactosylceramidase
galactosylceramidase I
-
-
-
-
galactosylceramide beta-galactosidase
-
-
-
-
galactosylcerebrosidase
-
-
-
-
galcerase
-
-
-
-
lactosylceramidase
-
-
-
-
lactosylceramidase I
-
-
-
-
beta-galactocerebrosidase
-
-
-
-
galactocerebrosidase
-
-
-
-
Galactosylceramidase
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of O-glycosyl bond
hydrolysis of O-glycosyl bond
-
-
-
-
PATHWAY SOURCE
PATHWAYS
SYSTEMATIC NAME
IUBMB Comments
D-galactosyl-N-acylsphingosine galactohydrolase
cf. EC 3.2.1.62 glycosylceramidase.
CAS REGISTRY NUMBER
COMMENTARY
9027-89-8
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
4-methylumbelliferyl beta-galactoside + H2O
methylumbelliferone + beta-D-galactose
-
-
-
-
?
4-methylumbelliferyl-beta-D-galactopyranoside + H2O
4-methylumbelliferone + beta-D-galactopyranose
-
pH 4.0, 30 min, 37°C
-
-
?
6-hexadecanoylamino-4-methylumbelliferyl-beta-D-galactopyranoside + H2O
6-hexadecanoylamino-4-methylumbelliferone + beta-D-galactopyranose
-
fluorogenic substrate
-
-
?
D-galactosyl-N-acylsphingosine + H2O
D-galactose + N-acylsphingosine
-
-
-
-
?
additional information
?
-
-
the enzyme is required for normal sperm maturation and function, enzyme deficiency leads to degeneration of oligodendrocytes, severe demyelination, and causes sperm abnormalities in the mouse model of human globoid cell leukodystrophy or Krabbe disease, mutant twitcher mice show reduced size of testis and sperm acrosomal membrane which is redundant, detached from the nucleus and folded over
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
D-galactosyl-N-acylsphingosine + H2O
D-galactose + N-acylsphingosine
-
-
-
-
?
additional information
?
-
-
the enzyme is required for normal sperm maturation and function, enzyme deficiency leads to degeneration of oligodendrocytes, severe demyelination, and causes sperm abnormalities in the mouse model of human globoid cell leukodystrophy or Krabbe disease, mutant twitcher mice show reduced size of testis and sperm acrosomal membrane which is redundant, detached from the nucleus and folded over
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.15
6-hexadecanoylamino-4-methylbelliferyl-beta-D-galactopyranoside
-
taurocholate and oleic acid activated
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.000008
-
GALC enzyme activity in the posterior part of the brain of single intracerebroventricular-GALC injected mice
0.0000087
-
GALC enzyme activity in the anterior part of the brain of single intracerebroventricular-GALC injected mice
120
-
with plasmid encoding GALC-MH transfected 293T cell, GALC units per mg of cell extract
130
-
with plasmid encoding unmodified GALC transfected 293T cell, GALC units per mg of cell extract
70
-
with plasmid encoding GALC-TMH transfected 293T cell, GALC units per mg of cell extract
additional information
additional information
-
economical and fast histochemical way to distinguish neural cells expressing galactocerebrosidase from thoses that are deficient, using 5Br-3Cl-beta-galactopiranoside in the presence of taurodeoxycholic and oleic acids
additional information
-
distribution of recombinant GALC activity in COS-7 cells: GALC-MH, 41.2% activity in cell extract, 58.8% released into cell culture medium, GALC-TMH, 27.5% to 72.5%
additional information
-
0.6 mU/mg for hematopoetic stem cells, after transduction with lentiviral GALC vector
additional information
-
0.65 mU/mg for hematopoetic stem cells from wild-type mice
additional information
-
2.33 mU/mg for neural precursor cells, after transduction with lentiviral GALC vector
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
3.5 - 5.3
-
pH 3.5: about 10% of maximal activity, pH 5.3: about 40% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
physiological function
-
lysosomal galactocerebrosidase GALC is involved in the maintenance of a functional hematopoietic stem/progenitor cell niche by contributing to the control of the intracellular content of key sphingolipids. Both insufficient and supraphysiologic GALC activity by inherited genetic deficiency or forced gene expression induce alterations of the intracellular content of the bioactive GALC downstream products ceramide and sphingosine, and thus affect hematopoietic stem/progenitor cell survival and function and the functionality of the stem cell niche
malfunction
heterozygote galactocerebrosidase mutants have reduced remyelination and impaired myelin debris clearance following demyelinating injury
malfunction
Krabbe's disease is a degenerative lysosomal storage disease resulting from deficiency of galactocerebrosidase activity. Over 100 mutations are known to cause the disease, and these usually occur in compound heterozygote patterns. In affected patients, nonsense mutations leading to a nonfunctional enzyme are often found associated with other mutations
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). GALC_MOUSE
684
1
77256
Swiss-Prot
Mitochondrion (Reliability: 4)
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystal structures of GALC and the GALC-D-galactose product complex, to 2.1 and 2.4 A resolution, respectively. The overall fold comprises a central triosephosphate isomerase barrel, a beta-sandwich domain, and a lectin domain. The overall fold of GALC is unchanged upon galactose binding, the core of the binding pocket being formed by the long loops on the C-terminal face of the TIM barrel. Loops from both the beta-sandwich and lectin domains also contribute to the substrate-binding pocket, and mutations involved in Krabbe's disease are widely distributed throughout the protein
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
E114K
residue in TIM barrel, mutation is likely to result in severe misfolding, crystallization data
E215K
residue is exposed on the surface of the TIM barrel, mutation confers an opposite charge on the same face as the substrate-binding pocket, crystallization data
G537R
resiude in lectin domain, mutation is likely to result in severe misfolding, crystallization data
L364R
resiude in beta-sandwich, mutation is likely to result in severe misfolding, crystallization data
L629R
resiude in lectin domain, mutation is likely to result in severe misfolding, crystallization data
S257F
resiude in TIM barrel, mutation is likely to result in severe misfolding, crystallization data
W339X
the twitcher mouse is a naturally occurring model of Krabbe's disease, containing in beta-galactocerebrosidase a premature stop codon, W339X
W410G
resiude in beta-sandwich, mutation is likely to result in severe misfolding, crystallization data
GALCdelta-MH
-
mutant containing a C-terminal myc-tag and six His residues with the last 11 amino acids at the C-terminus of GALC deleted
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression vectors for different fusion-proteins are constructed: GALC, unmodified enzyme, GALC-MH: fusion protein containing a C-terminal myc-tag and six His residues, GALC-TMH: fusion protein containing in addition a Tat-PTD, the protein transduction domain derived from the HIV-1 Tat protein
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
mutations involved in Krabbe's disease are widely distributed throughout the protein. Mutations that are likely to result in severe misfolding include E114K and S257F in the TIM barrel, L364R and W410G in the beta-sandwich domain, and G537R and L629R in the lectin domain. Mutation E215K is exposed on the surface of the TIM barrel. The mutation confers an opposite charge on the same face as the substrate-binding pocket suggesting that the mechanism of disease for this mutation will involve the perturbation of a binding face for an activating factor. Residue P302 that is found on the surface of GALC very close to the substrate-binding pocket is mutated to arginine in Krabbe's disease. The beta-sandwich domain of a long loop forms an integral part of the substrate-binding site. R380 at the tip of this loop directly binds the galactose molecule in the active site, its mutation to tryptophan or leucine leads to severe infantile Krabbe's disease
medicine
medicine
-
economical and fast histochemical way to distinguish neural cells expressing galactocerebrosidase from thoses that are deficient. This method enables the assessment of enzyme activity in virally-transduced cells as well as the biodistribution of galactocerebrosidase activity in Twitcher mice under gene or cell therapy
medicine
-
findings could lead to an improved therapy for globoid cell leukodystrophy, GLD, or Krabbe disease
medicine
-
globoid cell leukodystrophy, GLD or Krabbe disease, is caused by loss-of-fuction mutations in the GALC gene, injection of GALC improves the survival of the mouse model of GLD
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Kobayashi, T.; Shinnoh, N.; Kuroiwa, Y.
Metabolism of galactosylceramide in the twitcher mouse, an animal model of human globoid cell leukodystrophy
Biochim. Biophys. Acta
879
215-220
1986
Homo sapiens, Mus musculus
Rushton, A.R.; Dawson, G.
Glycosphinoglipid beta-galactosidases of cultured mammalian cells. Characterization of the enzymes from mouse cell line lmtk and human Lesch-Nyhan fibroblasts
Biochim. Biophys. Acta
388
92-105
1975
Homo sapiens, Mus musculus
Wiederschain, G.; Raghavan, S.; Kolodny, E.
Characterization of 6-hexadecanoylamino-4-methylumbelliferyl-beta-D-galactopyranoside as fluorogenic substrate of galactocerebrosidase for the diagnosis of Krabbe disease
Clin. Chim. Acta
205
87-96
1992
Homo sapiens, Mus musculus
Dolcetta, D.; Perani, L.; Givogri, M.I.; Galbiati, F.; Orlacchio, A.; Martino, S.; Roncarolo, M.G.; Bongarzone, E.
Analysis of galactocerebrosidase activity in the mouse brain by a new histological staining method
J. Neurosci. Res.
77
462-464
2004
Mus musculus
Luddi, A.; Strazza, M.; Carbone, M.; Moretti, E.; Costantino-Ceccarini, E.
Galactosylceramidase deficiency causes sperm abnormalities in the mouse model of globoid cell leukodystrophy
Exp. Cell Res.
304
59-68
2005
Mus musculus
Lee, W.C.; Tsoi, Y.K.; Troendle, F.J.; DeLucia, M.W.; Ahmed, Z.; Dicky, C.A.; Dickson, D.W.; Eckman, C.B.
Single-dose intracerebroventricular administration of galactocerebrosidase improves survival in a mouse model of globoid cell leukodystrophy
FASEB J.
21
2520-2527
2007
Mus musculus
Zhang, X.Y.; Dinh, A.; Cronin, J.; Li, S.C.; Reiser, J.
Cellular uptake and lysosomal delivery of galactocerebrosidase tagged with the HIV Tat protein transduction domain
J. Neurochem.
104
1055-1064
2008
Mus musculus
Martino, S.; Tiribuzi, R.; Tortori, A.; Conti, D.; Visigalli, I.; Lattanzi, A.; Biffi, A.; Gritti, A.; Orlacchio, A.
Specific determination of beta-galactocerebrosidase activity via competitive inhibition of beta-galactosidase
Clin. Chem.
55
541-548
2009
Homo sapiens, Mus musculus
Visigalli, I.; Ungari, S.; Martino, S.; Park, H.; Cesani, M.; Gentner, B.; Sergi Sergi, L.; Orlacchio, A.; Naldini, L.; Biffi, A.
The galactocerebrosidase enzyme contributes to the maintenance of a functional hematopoietic stem cell niche
Blood
116
1857-1866
2010
Homo sapiens, Mus musculus
Deane, J.E.; Graham, S.C.; Kim, N.N.; Stein, P.E.; McNair, R.; Cachon-Gonzalez, M.B.; Cox, T.M.; Read, R.J.
Insights into Krabbe disease from structures of galactocerebrosidase
Proc. Natl. Acad. Sci. USA
108
15169-15173
2011
Mus musculus (P54818)
Scott-Hewitt, N.J.; Folts, C.J.; Hogestyn, J.M.; Piester, G.; Mayer-Proeschel, M.; Noble, M.D.
Heterozygote galactocerebrosidase (GALC) mutants have reduced remyelination and impaired myelin debris clearance following demyelinating injury
Hum. Mol. Genet.
26
2825-2837
2017
Mus musculus (P54818)
Luddi, A.; Crifasi, L.; Capaldo, A.; Piomboni, P.; Costantino-Ceccarini, E.
Suppression of galactocerebrosidase premature termination codon and rescue of galactocerebrosidase activity in twitcher cells
J. Neurosci. Res.
94
1273-1283
2016
Homo sapiens (P54803), Homo sapiens, Mus musculus (P54818), Mus musculus
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