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Literature summary extracted from

  • Hayashi, Y.; Okino, N.; Kakuta, Y.; Shikanai, T.; Tani, M.; Narimatsu, H.; Ito, M.
    Klotho-related protein is a novel cytosolic neutral beta-glycosylceramidase (2007), J. Biol. Chem., 282, 30889-30900.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.2.1.62 sodium cholate increases the solubility of the substrates Homo sapiens

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.45 expressed in Escherichia coli BL21(DE3)pLysS cells Homo sapiens
3.2.1.62 expression in Chinese hamster ovary-derived cells expressing polyoma LT antigen Mus musculus
3.2.1.62 expression in Chinese hamster ovary-derived cells expressing polyoma LT antigen, in HEK293 cells cotransfected with in KlrP siRNA, overexpression in Escherichia coli Homo sapiens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.45 hanging drop vapour diffusion method with 0.1 M Tris-HCl buffer, pH 8.5, containing 0.2 M magnesium chloride, 27.5% PEG 3350, 5% glycerol, 0.5 M glucose or 0.5 M galactose Homo sapiens
3.2.1.62 of the recombinant protein with the hanging drops by vapor diffusion method Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.62 E165D reduced activity Homo sapiens
3.2.1.62 E165Q almost no activity Homo sapiens
3.2.1.62 E373D reduced activity Homo sapiens
3.2.1.62 E373Q almost no activity Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.62 additional information conduritol B epoxide only slightly inhibitory Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.45 0.00204
-
 (6-[N-(7-nitrobenz-2-oxa-1,3 diazol-4-yl)amino]hexanoyl)-beta-D-galactosylceramide in 50 mM MES buffer, pH 6.0, at 37°C Homo sapiens
3.2.1.45 0.00464
-
 (6-[N-(7-nitrobenz-2-oxa-1,3 diazol-4-yl)amino]hexanoyl)-beta-D-glucosylceramide in 50 mM MES buffer, pH 6.0, at 37°C Homo sapiens
3.2.1.45 0.00924
-
 C18-galactosylceramide in 50 mM MES buffer, pH 6.0, at 37°C Homo sapiens
3.2.1.45 0.01367
-
 C18-glucosylceramide in 50 mM MES buffer, pH 6.0, at 37°C Homo sapiens
3.2.1.45 0.04928
-
 4-methylumbelliferyl-beta-D-glucopyranoside in 50 mM MES buffer, pH 6.0, at 37°C Homo sapiens
3.2.1.45 0.1445
-
 4-methylumbelliferyl-beta-D-galactopyranoside in 50 mM MES buffer, pH 6.0, at 37°C Homo sapiens
3.2.1.62 2.04
-
 C6-4-nitrobenzo-2-oxa-1,3-diazole-galctosylceramide
-
 Homo sapiens
3.2.1.62 4.64
-
 (6-[N-(7-nitrobenz-2-oxa-1,3 diazol-4-yl)amino]hexanoyl)-beta-D-glucosylceramide
-
 Homo sapiens
3.2.1.62 9.24
-
 C18-galactosylceramide
-
 Homo sapiens
3.2.1.62 13.67
-
 C18-glucosylceramide
-
 Homo sapiens
3.2.1.62 49.28
-
 4-methylumbelliferyl-beta-D-glucopyranoside
-
 Homo sapiens
3.2.1.62 144.5
-
 4-methylumbelliferyl-beta-D-galactopyranoside
-
 Homo sapiens

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.2.1.45 cytosol
-
 Homo sapiens 5829
-
3.2.1.62 cytosol
-
 Mus musculus 5829
-
3.2.1.62 cytosol
-
 Homo sapiens 5829
-
3.2.1.62 cytosol
-
 Rattus norvegicus 5829
-
3.2.1.62 cytosol
-
 Danio rerio 5829
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.62 57000
-
 SDS-PAGE Homo sapiens

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.45 Homo sapiens
-
-
-
3.2.1.62 Danio rerio
-
-
-
3.2.1.62 Homo sapiens
-
-
-
3.2.1.62 Mus musculus
-
-
-
3.2.1.62 Rattus norvegicus
-
-
-
3.2.1.62 Tetraodontidae
-
 puffer fish, no activity in Mycetozoa, slime mold
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.45 Hi-Trap Chelating HP column chromatography and Superdex 200 10/300 gel filtration Homo sapiens
3.2.1.62 of the recombinant protein by a hi-trap chelating HP column and superdex 200 column chromatography Homo sapiens

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.2.1.45 fibroblast
-
 Homo sapiens
-
3.2.1.62 brain
-
 Rattus norvegicus
-
3.2.1.62 embryo
-
 Danio rerio
-
3.2.1.62 fibroblast
-
 Homo sapiens
-
3.2.1.62 liver
-
 Mus musculus
-
3.2.1.62 liver
-
 Tetraodontidae
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.62 0.00016
-
 activity in CHOP cell lysates transformed with cDNA encoding KLPH, with conduritol B epoxide, C6-4-nitrobenzo-2-oxa-1,3-diazole-galactosylceramide as substrate Mus musculus
3.2.1.62 0.00025
-
 activity in CHOP cell lysates transformed with cDNA encoding KLPH, without conduritol B epoxide, C6-4-nitrobenzo-2-oxa-1,3-diazole-galactosylceramide as substrate Mus musculus
3.2.1.62 0.0005
-
 activity in CHOP cell lysates transformed with cDNA encoding KLPH, with conduritol B epoxide, C6-4-nitrobenzo-2-oxa-1,3-diazole-glucosylceramide as substrate Mus musculus
3.2.1.62 0.0005
-
 in membrane fraction with conduritol B epoxide, C6-4-nitrobenzo-2-oxa-1,3-diazole-glucosylceramide as substrate Rattus norvegicus
3.2.1.62 0.00133
-
 in cytosolic fraction with conduritol B epoxide, C6-4-nitrobenzo-2-oxa-1,3-diazole-glucosylceramide as substrate Rattus norvegicus
3.2.1.62 0.001875
-
 activity in CHOP cell lysates transformed with cDNA encoding KlrP, with conduritol B epoxide, C6-4-nitrobenzo-2-oxa-1,3-diazole-glucosylceramide as substrate Homo sapiens
3.2.1.62 0.0025
-
 activity in CHOP cell lysates transformed with cDNA encoding KLPH, without conduritol B epoxide, C6-4-nitrobenzo-2-oxa-1,3-diazole-glucosylceramide as substrate Mus musculus
3.2.1.62 0.00375
-
 activity in CHOP cell lysates transformed with cDNA encoding KlrP, without conduritol B epoxide, C6-4-nitrobenzo-2-oxa-1,3-diazole-glucosylceramide as substrate Homo sapiens
3.2.1.62 0.0052
-
 activity in CHOP cell lysates transformed with cDNA encoding KlrP, with conduritol B epoxide, C6-4-nitrobenzo-2-oxa-1,3-diazole-galactosylceramide as substrate Homo sapiens
3.2.1.62 0.0066
-
 activity in CHOP cell lysates transformed with cDNA encoding KlrP, without conduritol B epoxide, C6-4-nitrobenzo-2-oxa-1,3-diazole-galactosylceramide as substrate Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.45 (6-[N-(7-nitrobenz-2-oxa-1,3 diazol-4-yl)amino]hexanoyl)-beta-D-glucosylceramide + H2O
-
 Homo sapiens (6-[N-(7-nitrobenz-2-oxa-1,3 diazol-4-yl)amino]hexanoyl)sphingosine + D-glucose
-
 ?
3.2.1.45 4-methylumbelliferyl-beta-D-galactopyranoside + H2O
-
 Homo sapiens methylumbelliferone + beta-D-galactose
-
 ?
3.2.1.45 4-methylumbelliferyl-beta-D-glucopyranoside + H2O
-
 Homo sapiens methylumbelliferone + beta-D-glucose
-
 ?
3.2.1.45 6-[N-(7-nitrobenz-2-oxa-1,3 diazol-4-yl)amino]hexanoyl)-beta-D-galactosylceramide + H2O
-
 Homo sapiens 6-[N-(7-nitrobenz-2-oxa-1,3 diazol-4-yl)amino]hexanoyl)ceramide + D-galactose
-
 ?
3.2.1.45 C18-galactosylceramide + H2O
-
 Homo sapiens ?
-
 ?
3.2.1.45 C18-glucosylceramide + H2O
-
 Homo sapiens ?
-
 ?
3.2.1.45 glucocerebroside + H2O
-
 Homo sapiens beta-D-glucose + ceramide
-
 ?
3.2.1.62 4-methylumbelliferyl-beta-D-galactopyranoside + H2O
-
 Homo sapiens 4-methylumbelliferone + D-galactose
-
 ?
3.2.1.62 4-methylumbelliferyl-beta-D-glucopyranoside + H2O
-
 Homo sapiens 4-methylumbelliferone + D-glucose
-
 ?
3.2.1.62 C18-galactosylceramide + H2O
-
 Homo sapiens C18-ceramide + D-galactose
-
 ?
3.2.1.62 C18-glucosylceramide + H2O
-
 Homo sapiens C18-ceramide + D-glucose
-
 ?
3.2.1.62 C6-4-nitrobenzo-2-oxa-1,3-diazole-galactosylceramide + H2O
-
 Mus musculus C6-4-nitrobenzo-2-oxa-1,3-diazole-ceramide + D-galactose
-
 ?
3.2.1.62 C6-4-nitrobenzo-2-oxa-1,3-diazole-galactosylceramide + H2O
-
 Homo sapiens C6-4-nitrobenzo-2-oxa-1,3-diazole-ceramide + D-galactose
-
 ?
3.2.1.62 C6-4-nitrobenzo-2-oxa-1,3-diazole-glucosylceramide + H2O C6-NBD-glucosylceramide Mus musculus C6-4-nitrobenzo-2-oxa-1,3-diazole-ceramide + D-glucose
-
 ?
3.2.1.62 C6-4-nitrobenzo-2-oxa-1,3-diazole-glucosylceramide + H2O C6-NBD-glucosylceramide Homo sapiens C6-4-nitrobenzo-2-oxa-1,3-diazole-ceramide + D-glucose
-
 ?
3.2.1.62 C6-4-nitrobenzo-2-oxa-1,3-diazole-glucosylceramide + H2O C6-NBD-glucosylceramide Rattus norvegicus C6-4-nitrobenzo-2-oxa-1,3-diazole-ceramide + D-glucose
-
 ?
3.2.1.62 C6-4-nitrobenzo-2-oxa-1,3-diazole-glucosylceramide + H2O C6-NBD-glucosylceramide Danio rerio C6-4-nitrobenzo-2-oxa-1,3-diazole-ceramide + D-glucose
-
 ?
3.2.1.62 C6-4-nitrobenzo-2-oxa-1,3-diazole-glucosylceramide + H2O C6-NBD-glucosylceramide Tetraodontidae C6-4-nitrobenzo-2-oxa-1,3-diazole-ceramide + D-glucose
-
 ?
3.2.1.62 galactosylceramide + H2O
-
 Homo sapiens ceramide + D-galactose
-
 ?
3.2.1.62 galactosylsphingosine + H2O
-
 Homo sapiens sphingosine + D-galactose
-
 ?
3.2.1.62 glucosylceramide + H2O no activity with alpha-galactosylceramide, lactosylceramide, ganglioside M1a and sulfatides Homo sapiens ceramide + D-glucose
-
 ?
3.2.1.62 glucosylsphingosine + H2O
-
 Homo sapiens beta-D-glucose + N-acylsphingosine
-
 ?
3.2.1.62 glycosyl-N-acylsphingosine + H2O
-
 Mus musculus N-acylsphingosine + a sugar
-
 ?
3.2.1.62 glycosyl-N-acylsphingosine + H2O
-
 Danio rerio N-acylsphingosine + a sugar
-
 ?
3.2.1.62 glycosyl-N-acylsphingosine + H2O
-
 Tetraodontidae N-acylsphingosine + a sugar
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.45 beta-glycosylceramidase
-
 Homo sapiens
3.2.1.45 Gcase
-
 Homo sapiens
3.2.1.45 Klotho-related protein
-
 Homo sapiens
3.2.1.45 KLrP
-
 Homo sapiens
3.2.1.45 neutral beta-glycosylceramidase
-
 Homo sapiens
3.2.1.62 Gcase
-
 Homo sapiens
3.2.1.62 Gcase
-
 Rattus norvegicus
3.2.1.62 Gcase
-
 Danio rerio
3.2.1.62 Gcase
-
 Tetraodontidae
3.2.1.62 glycosyl-N-acylsphingosine glycohydrolase
-
 Mus musculus
3.2.1.62 glycosyl-N-acylsphingosine glycohydrolase
-
 Homo sapiens
3.2.1.62 glycosyl-N-acylsphingosine glycohydrolase
-
 Rattus norvegicus
3.2.1.62 glycosyl-N-acylsphingosine glycohydrolase
-
 Danio rerio
3.2.1.62 glycosyl-N-acylsphingosine glycohydrolase
-
 Tetraodontidae
3.2.1.62 glycosylceramidase
-
 Mus musculus
3.2.1.62 glycosylceramidase
-
 Homo sapiens
3.2.1.62 glycosylceramidase
-
 Rattus norvegicus
3.2.1.62 glycosylceramidase
-
 Danio rerio
3.2.1.62 glycosylceramidase
-
 Tetraodontidae
3.2.1.62 KLotho lactase phlorizin hydrolase KLPH Mus musculus
3.2.1.62 Klotho-related protein KlrP, novel cytosolic neutral glycosylceramidase Homo sapiens
3.2.1.62 Klotho-related protein KlrP, novel cytosolic neutral glycosylceramidase Rattus norvegicus
3.2.1.62 Klotho-related protein KlrP, novel cytosolic neutral glycosylceramidase Danio rerio
3.2.1.62 Klotho-related protein KlrP, novel cytosolic neutral glycosylceramidase Tetraodontidae

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.45 0.007
-
 C18-glucosylceramide in 50 mM MES buffer, pH 6.0, at 37°C Homo sapiens
3.2.1.45 0.03
-
 (6-[N-(7-nitrobenz-2-oxa-1,3 diazol-4-yl)amino]hexanoyl)-beta-D-galactosylceramide in 50 mM MES buffer, pH 6.0, at 37°C Homo sapiens
3.2.1.45 0.12
-
 (6-[N-(7-nitrobenz-2-oxa-1,3 diazol-4-yl)amino]hexanoyl)-beta-D-glucosylceramide in 50 mM MES buffer, pH 6.0, at 37°C Homo sapiens
3.2.1.45 1.23
-
 4-methylumbelliferyl-beta-D-glucopyranoside in 50 mM MES buffer, pH 6.0, at 37°C Homo sapiens
3.2.1.45 1.57
-
 4-methylumbelliferyl-beta-D-galactopyranoside in 50 mM MES buffer, pH 6.0, at 37°C Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.45 5
-
-
 Homo sapiens
3.2.1.62 6 7 C6-4-nitrobenzo-2-oxa-1,3-diazole-glucosylceramide as substrate Homo sapiens

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.62 5 8
-
 Homo sapiens