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Literature summary for 2.4.1.41 extracted from

  • Tenno, M.; Saeki, A.; Elhammer, A.P.; Kurosaka, A.
    Function of conserved aromatic residues in the Gal/GalNAc-glycosyltransferase motif of UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase 1 (2007), FEBS J., 274, 6037-6045.
    View publication on PubMed
Search for more literature for 2.4.1.41

Application

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(Commentary)

Cloned (Comment) Organism
expression in COS7 cells as truncated version from pInsProA Rattus norvegicus

Protein Variants

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 [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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Rattus norvegicus Q10473 GalNAc-T1
-

Purification (Commentary)

Purification (Comment) Organism
recovery of the secreted recombinant proteins that harbour a protein A IgG-binding domain by IgG-Sepharose Rattus norvegicus

Substrates and Products (Substrate)

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

Synonyms Comment Organism
GalNAc transferase
-
 Rattus norvegicus
GalNAc-T
-
 Rattus norvegicus
UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase
-
 Rattus norvegicus