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Literature summary for 2.4.1.101 extracted from

  • Saribas, A.S.; Johnson, K.; Liu, L.; Bezila, D.; Hakes, D.
    Refolding of human beta-1-2 GlcNAc transferase (GnT1) and the role of its unpaired Cys121 (2007), Biochem. Biophys. Res. Commun., 362, 381-386.
    View publication on PubMed
Search for more literature for 2.4.1.101

Cloned(Commentary)

Cloned (Comment) Organism
human beta-1-2-N-acetylglucosaminyltransferase (hGnT1) lacking the first 103 amino acids is expressed as a maltose binding protein (MBP) fusion protein in inclusion bodies in Escherichia coli and refolded using an oxido-shuffling method. Cloning and expression of MBP-hGnT1(D103) and mutant MBP-hGnT1 (D103) enzymes Homo sapiens

Protein Variants

Protein Variants Comment Organism
C121A active mutant enzyme. After 1 h, the wild-type MBP-hGnT1(D103) and C121A mutant convert more than 50% of Man5-RNAse B to RNAse B-GlcNAc2-Man5-GlcNAc Homo sapiens
C121D mutant enzyme shows no transfer of GlcNAc from UDP-GlcNAc to the glycoprotein acceptor Man5-RNAse B Homo sapiens
C121S active mutant enzyme. After 1 h, the C121S mutant shows less than 25% conversion of Man5-RNAse B to RNAse B-GlcNAc2-Man5-GlcNAc Homo sapiens
C121T mutant enzyme shows no transfer of GlcNAc from UDP-GlcNAc to the glycoprotein acceptor Man5-RNAse B Homo sapiens
R120A/C121H active mutant enzyme. The R120A/C121H mutant is less active than the wild type and the single mutants. It converts more than half of the Man5-RNAse B to RNAse B-GlcNAc2-Man5-GlcNAc Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
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Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
UDP-N-acetyl-D-glucosamine + Man5-RNAse B wild type hGnT1 and mutants C121A, C121S and R120A/C121H transfer GlcNAc from UDP-GlcNAc to the glycoprotein acceptor Man5-RNAse B, whereas mutants C121T and C121D are inactive. After 1 h, the wild-type MBP-hGnT1(D103) and C121A mutant convert more than 50% of RNAse BM5 to RNAse BM5Gn, while the C121S mutant shows less than 25% conversion. The wild-type MBP-hGnT1(D103) and both C121A and C121S mutants convert RNAse BM5 nearly completely to RNAse BM5Gn following overnight incubation. The R120A/C121H mutant is less active than the wild-type and the single mutants. It converts more than half of the RNAse BM5 into RNAse BM5Gn after 17 h Homo sapiens UDP + RNAse B-GlcNAc2-Man5-GlcNAc
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Synonyms

Synonyms Comment Organism
beta-1-2 GlcNAc transferase
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 Homo sapiens
GnT1
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 Homo sapiens