Application | Comment | Organism |
---|---|---|
molecular biology | invariant residue Trp328 is essential for GalNAc-T enzymatic activity, residue Trp316 is important in the interaction with the acceptor polypeptide of GalNAc-T1 | Rattus norvegicus |
Cloned (Comment) | Organism |
---|---|
expression in COS7 cells as truncated version from pInsProA | Rattus norvegicus |
Protein Variants | Comment | Organism |
---|---|---|
F303L | mutant of 40% activity relative to truncated wild-type using apomucin as acceptor substrate and of increased KM for both substrates | Rattus norvegicus |
F325L | mutant of 60% activity relative to truncated wild-type using apomucin as acceptor substrate | Rattus norvegicus |
additional information | all point mutants are mutants of the truncated wild-type P-T1-delta42 and were generated by site-directed mutagenesis on pInsProAdelta42 | Rattus norvegicus |
P-T1-delta42 | truncated GalNAc-T1 wild-type, deletion of the coding sequence for the cytoplasmic tail and the transmembrane domain, cDNA for insulin signal sequence and protein A IgG-binding domain fused to the 5ย-end | Rattus norvegicus |
W316A | inactive mutant using apomucin as acceptor substrate | Rattus norvegicus |
W316F | mutant of 20% activity relative to truncated wild-type using apomucin as acceptor substrate and of increased KMs for apomucin, UDP-GalNAc, and synthetic peptides, PPDAATAAPL and GVVPTVVPG | Rattus norvegicus |
W316L | background level activity relative to truncated wild-type using apomucin as acceptor substrate | Rattus norvegicus |
W316Y | mutant of 40% activity relative to truncated wild-type using apomucin as acceptor substrate and of increased KMs for apomucin, UDP-GalNAc, and synthetic peptides, PPDAATAAPL and GVVPTVVPG | Rattus norvegicus |
W328A | inactive mutant using apomucin as acceptor substrate, low expression level | Rattus norvegicus |
W328F | inactive mutant using apomucin as acceptor substrate | Rattus norvegicus |
W328L | inactive mutant using apomucin as acceptor substrate | Rattus norvegicus |
W328Y | inactive mutant using apomucin as acceptor substrate | Rattus norvegicus |
Y302L | mutant of 80% activity relative to truncated wild-type using apomucin as acceptor substrate | Rattus norvegicus |
Y309L | mutant of 40% activity relative to truncated wild-type using apomucin as acceptor substrate and of increased KM for both substrates | Rattus norvegicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
apomucin | 13.6 +/-1.9 mg/ml, mutant F303L, 2.8 times higher than wild-type | Rattus norvegicus | |
additional information | - |
apomucin | 17.4 +/-3.6 mg/ml, mutant W316Y, 3.6 times higher than wild-type | Rattus norvegicus | |
additional information | - |
apomucin | 4.7 +/-0.1 mg/ml, truncated wild-type P-T1/delta42 | Rattus norvegicus | |
additional information | - |
apomucin | 43.9 +/-3.8 mg/ml, mutant W316F, 9.1 times higher than wild-type | Rattus norvegicus | |
additional information | - |
apomucin | 5.0 +/-0.2 mg/ml, mutant Y302L | Rattus norvegicus | |
additional information | - |
apomucin | 6.7 +/-1.7 mg/ml, mutant F325L, 1.4 times higher than wild-type | Rattus norvegicus | |
additional information | - |
apomucin | 7.1 +/-0.4 mg/ml, mutant Y309L, 1.5 times higher than wild-type | Rattus norvegicus | |
additional information | - |
additional information | KMs obtained using Lineweaver-Burk plot | Rattus norvegicus | |
additional information | - |
additional information | the KMs for apomucin and the synthetic peptides, PPDAATAAPL and GVVPTVVPG, were measured in presence of 7.5 ยตM UDP-GalNAc | Rattus norvegicus | |
additional information | - |
additional information | the KMs for UDP-GalNAc were measured in presence of 1.88 mg/ml apomucin | Rattus norvegicus | |
0.35 | - |
PPDAATAAPL | +/-0.05 mM, truncated wild-type P-T1/delta42 | Rattus norvegicus | |
1.74 | - |
GVVPTVVPG | +/-0.3 mM, truncated wild-type P-T1/delta42 | Rattus norvegicus | |
5.1 | - |
UDP-GalNAc | +/-0.8 mM, truncated wild-type P-T1/delta42 | Rattus norvegicus | |
5.2 | - |
UDP-GalNAc | +/-0.1 mM, mutant Y302L | Rattus norvegicus | |
6.2 | - |
UDP-GalNAc | +/-0.6 mM, mutant F325L, 1.2 times higher than wild-type | Rattus norvegicus | |
9 | - |
UDP-GalNAc | +/-0.1 mM, mutant W316Y, 1.8 times higher than wild-type | Rattus norvegicus | |
9.1 | - |
UDP-GalNAc | +/-0.7 mM, mutant W316F, 1.8 times higher than wild-type | Rattus norvegicus | |
13.5 | - |
PPDAATAAPL | +/-0.7 mM, mutant W316Y, 38.5 times higher than wild-type | Rattus norvegicus | |
14.2 | - |
UDP-GalNAc | +/-1.1 mM, mutant F303L, 2.8 times higher than wild-type | Rattus norvegicus | |
15.6 | - |
PPDAATAAPL | +/-0.6 mM, mutant W316F, 44.6 times higher than wild-type | Rattus norvegicus | |
16 | - |
GVVPTVVPG | +/-0.3 mM, mutant W316Y, 9.2 times higher than wild-type | Rattus norvegicus | |
16 | - |
UDP-GalNAc | +/-0.3 mM, mutant Y309L, 3.2 times higher than wild-type | Rattus norvegicus | |
17.6 | - |
GVVPTVVPG | +/-0.5 mM, mutant W316F, 10.1 times higher than wild-type | Rattus norvegicus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-N-acetyl-D-galactosamine + polypeptide | Rattus norvegicus | mucin-type O-linked glycosylation of serine or threonine residues | UDP + N-acetyl-D-galactosaminyl-polypeptide | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rattus norvegicus | Q10473 | GalNAc-T1 | - |
Purification (Comment) | Organism |
---|---|
recovery of the secreted recombinant proteins that harbour a protein A IgG-binding domain by IgG-Sepharose | Rattus norvegicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-GalNAc + apomucin | apomucin is a deglycosylated bovine submaxillary mucin, mutants W328Y, W328F, W328A, W316H and W316A are inactive, mutants W316Y and W316F show 40% and 20% activity, respectively, relative to wild-type | Rattus norvegicus | UDP + GalNAc-mucin | - |
? | |
UDP-GalNAc + GVVPTVVPG | GVVPTVVPG is a synthetic peptide with threonine as acceptor site, weaker acceptor substrate for wild-type and truncated version P-T1/delta42 as PPDAATAAPL | Rattus norvegicus | UDP + GVVP(GalNAc)TVVPG | - |
? | |
UDP-GalNAc + PPDAATAAPL | PPDAATAAPL is a synthetic peptide with threonine as acceptor site, efficient acceptor substrate for wild-type and truncated version P-T1/delta42 | Rattus norvegicus | UDP + PPDAA(GalNAc)TAAPL | - |
? | |
UDP-N-acetyl-D-galactosamine + polypeptide | mucin-type O-linked glycosylation of serine or threonine residues | Rattus norvegicus | UDP + N-acetyl-D-galactosaminyl-polypeptide | - |
? |
Synonyms | Comment | Organism |
---|---|---|
GalNAc transferase | - |
Rattus norvegicus |
GalNAc-T | - |
Rattus norvegicus |
UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase | - |
Rattus norvegicus |