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Literature summary extracted from

  • Marks, D.L.; Dominguez, M.; Wu, K.; Pagano, R.E.
    Identification of active site residues in glucosylceramide synthase. A nucleotide-binding/catalytic motif conserved with processive beta-glycosyltransferases (2001), J. Biol. Chem., 276, 26492-26498.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.80 expression of wild-type and mutants in Escherichia coli BL21(DE3) Rattus norvegicus

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.80 C143A site-directed mutagenesis, increased activity, reduced expression level compared to wild-type Rattus norvegicus
2.4.1.80 C207A site-directed mutagenesis, reduced activity, reduced inhibitory effect of N-ethylmaleimide, reduced expression level compared to wild-type Rattus norvegicus
2.4.1.80 C213A site-directed mutagenesis, reduced activity, reduced expression level compared to wild-type Rattus norvegicus
2.4.1.80 C232A site-directed mutagenesis, reduced expresssion level compared to wild-type Rattus norvegicus
2.4.1.80 C290A site-directed mutagenesis, reduced expression level compared to wild-type Rattus norvegicus
2.4.1.80 C296A site-directed mutagenesis, reduced activity, reduced expression level compared to wild-type Rattus norvegicus
2.4.1.80 C321A site-directed mutagenesis, reduced activity Rattus norvegicus
2.4.1.80 C321A/C323A site-directed mutagenesis, reduced activity Rattus norvegicus
2.4.1.80 C343A site-directed mutagenesis, enhanced activity, slightly reduced expression level compared to wild-type Rattus norvegicus
2.4.1.80 C384A site-directed mutagenesis, enhanced activity Rattus norvegicus
2.4.1.80 C86A site-directed mutagenesis, reduced activity, reduced expression level compared to wild-type Rattus norvegicus
2.4.1.80 C98A site-directed mutagenesis, reduced activity, reduced expression level compared to wild-type Rattus norvegicus

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.4.1.80 N-ethylmaleimide attacks Cys207 Rattus norvegicus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.1.80 additional information
-
 amino acid sequence of conserved D1,D2,D3,QXXRW motif Rattus norvegicus

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.80 Rattus norvegicus Q9R0E0
-
-

Reaction

EC Number Reaction Comment Organism Reaction ID
2.4.1.80 UDP-alpha-D-glucose + an N-acylsphingosine = UDP + a beta-D-glucosyl-N-acylsphingosine UDP-glucose and inhibitor D-PDMP binding region overlap and contain His193 Rattus norvegicus
a
2.4.1.80 UDP-alpha-D-glucose + an N-acylsphingosine = UDP + a beta-D-glucosyl-N-acylsphingosine enzyme DNA sequence contains conserved D1,D2,D3,QXXRW motif found also in other progressive beta-glycosyltransferases, putative active site, Cys207 and His193 involved Rattus norvegicus
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.80 UDP-glucose + N-acylsphingosine i.e. ceramide Rattus norvegicus UDP + D-glucosyl-N-acylsphingosine i.e. glucosylceramide ?
2.4.1.80 UDP-glucose + N-[7-(4-nitrobenzo-2-oxa-1,3-diazole)]-6-aminocaproyl-D-erythro-sphingosine synthetic fluorescent ceramide substrate analogue for fluorescence enzyme assay Rattus norvegicus UDP + D-glucosyl-N-[7-(4-nitrobenzo-2-oxa-1,3-diazole)]-6-aminocaproyl-D-erythro-sphingosine
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.4.1.80 GCS
-
 Rattus norvegicus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.1.80 37
-
 assay at Rattus norvegicus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.80 7.4
-
 assay at Rattus norvegicus