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Literature summary for 2.4.1.229 extracted from

  • van der Wel, H.; Morris, H.R.; Panico, M.; Paxton, T.; Dell, A.; Kaplan, L.; West, C.M.
    Molecular cloning and expression of a UDP-N-acetylglucosamine (GlcNAc):hydroxyproline polypeptide GlcNAc-transferase that modifies Skp1 in the cytoplasm of Dictyostelium (2002), J. Biol. Chem., 277, 46328-46337.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene GnT51, DNA sequence determination and analysis, functional expression of wild-type and mutants as fusion proteins with CBD-intein, in Escherichia coli strain ER2566, autocleavage of tag moiety Dictyostelium discoideum

Protein Variants

Protein Variants Comment Organism
D102A site-directed mutagenesis, no remaining activity Dictyostelium discoideum
H104D site-directed mutagenesis, no remaining activity Dictyostelium discoideum
K23R site-directed mutagenesis, unaltered enzyme activity Dictyostelium discoideum
L276S site-directed mutagenesis, 7% remaining activity compared to the wild-type enzyme Dictyostelium discoideum
additional information C-terminal deletion mutants GnT51(1-282) and GnT51(1-369+) show no remaining activity Dictyostelium discoideum

Localization

Localization Comment Organism GeneOntology No. Textmining
cytosol
-
Dictyostelium discoideum 5829
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
29600
-
x * 29600, mutant GnT51(1-282), SDS-PAGE Dictyostelium discoideum
44000
-
x * 44000, mutant GnT51(1-369+), SDS-PAGE Dictyostelium discoideum
47500
-
x * 47500, wild-type enzyme and mutants K23R, D102A, H104D, and L276S, SDS-PAGE Dictyostelium discoideum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
UDP-N-acetylglucosamine + [Skp1-protein]-hydroxyproline Dictyostelium discoideum
-
UDP + [Skp1-protein]-O-(N-acetyl-D-glucosaminyl)hydroxyproline
-
?

Organism

Organism UniProt Comment Textmining
Dictyostelium discoideum Q8T1C6 gene GnT51
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type enzyme and mutants from Escherichia coli as CBD-intein tagged proteins, to near homogeneity Dictyostelium discoideum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
UDP-N-acetylglucosamine + [Skp1-protein]-hydroxyproline
-
Dictyostelium discoideum UDP + [Skp1-protein]-O-(N-acetyl-D-glucosaminyl)hydroxyproline
-
?
UDP-N-acetylglucosamine + [Skp1-protein]-hydroxyproline enzyme attaches the first sugar of pentasaccharide chain Dictyostelium discoideum UDP + [Skp1-protein]-O-(N-acetyl-D-glucosaminyl)hydroxyproline
-
?
UDP-N-acetylglucosamine + [Skp1-protein]-hydroxyproline hydroxyproline143 Dictyostelium discoideum UDP + [Skp1-protein]-O-(N-acetyl-D-glucosaminyl)hydroxyproline
-
?

Subunits

Subunits Comment Organism
? x * 47500, wild-type enzyme and mutants K23R, D102A, H104D, and L276S, SDS-PAGE Dictyostelium discoideum
? x * 44000, mutant GnT51(1-369+), SDS-PAGE Dictyostelium discoideum
? x * 29600, mutant GnT51(1-282), SDS-PAGE Dictyostelium discoideum

Synonyms

Synonyms Comment Organism
GnT51
-
Dictyostelium discoideum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.8
-
assay at Dictyostelium discoideum