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Literature summary for 2.4.1.41 extracted from

  • Raman, J.; Fritz, T.A.; Gerken, T.A.; Jamison, O.; Live, D.; Liu, M.; Tabak, L.A.
    The catalytic and lectin domains of UDP-GalNAc:polypeptide alpha-N-acetylgalactosaminyltransferase function in concert to direct glycosylation site selection (2008), J. Biol. Chem., 283, 22942-22951.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.4.1.41

Cloned(Commentary)

Cloned (Comment) Organism
in pKN55-N6His-TEV for expression in Pichia pastoris strain SMD1168 and with TEV-cleavable hexa-His-tag Homo sapiens

Protein Variants

Protein Variants Comment Organism
hT10 GalNAc-T10, lacking transmembrane domain (AA1-70) Homo sapiens
hT10CD GalNAc-T10 catalytic domain, hT10 lacking lectin domain (AA446-603) and transmembrane domain (AA1-70), no altered preference in glycosylation site selection, loss of initial burst phase during glycosylation of MUC5AC-13 at Thr-9, gain of initial burst phase during glycosylation of MUC5AC-3 at Thr-9, GalNAc-transfer accompanied by high rate of UDP-GalNAc hydrolysis, lectin domain not required for catalysis, no glycosylation of MUC5AC-9 Homo sapiens
hT10CD-hT2LD GalNAc-T10 catalytic domain-GalNAc-T2 lectin domain, hT10 lacking lectin domain (AA446-603) and transmembrane domain (AA1-70) fused to hT2 lectin domain, no glycosylation of MUC5AC-9 Homo sapiens
hT2 GalNAc-T2, lacking transmembrane domain (AA1-74) Homo sapiens
hT2CD GalNAc-T2 catalytic domain, hT2 lacking lectin domain (AA441-571) and transmembrane domain (AA1-74), reduced affinity for substrate MUC5AC-13 but not for MUC5AC-3, shift of preferred MUC5AC-3 glycosylation site from Thr-9 to Thr-13 Homo sapiens
hT2CD-hT10LD GalNAc-T2 catalytic domain-GalNAc-T10 lectin domain, hT2 lacking lectin domain (AA441-571) and transmembrane domain (AA1-74) fused to hT10 lectin domain, partially restored glycosylation to Thr-3 on MUC5AC-13 but not to Thr-13 on MUC5AC-3 or Ser-5 on MUC5AC-3,13 Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.018
-
 MUC5AC-13 at site Thr-3, hT2 Homo sapiens
0.018
-
 MUC5AC at site Thr-9, hT2CD Homo sapiens
0.02
-
 MUC5AC at site Thr-9, hT2 Homo sapiens
0.033
-
 MUC5AC-3 at site Thr-13, hT2 Homo sapiens
0.036
-
 MUC5AC-3 at site Thr-9, hT2CD Homo sapiens
0.05
-
 MUC5AC-3 at site Thr-2, hT10CD Homo sapiens
0.096
-
 MUC5AC-3 at site Thr-2, hT10 Homo sapiens
0.107
-
 MUC5AC-3 at site Thr-2, hT10CD-hT2LD Homo sapiens
0.17
-
 MUC5AC-13 at site Thr-9, hT2CD Homo sapiens
0.44
-
 MUC5AC-3,13 at sites Thr-2 and Thr12, hT10 Homo sapiens
0.53
-
 MUC5AC-13 at site Thr-12, hT10 Homo sapiens
0.57
-
 MUC5AC-3,13 at sites Thr-2 and Thr12, hT10CD-hT2LD Homo sapiens
0.64
-
 MUC5AC-13 at site Thr-12, hT10CD-hT2LD Homo sapiens
0.77
-
 MUC5AC-13 at site Thr-12, hT10CD Homo sapiens
1
-
 MUC5AC-3,13 at sites Thr-2 and Thr12, hT10CD Homo sapiens
2.4
-
 MUC5AC-9 at site Thr-2, hT10 Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
UDP-N-acetyl-D-galactosamine + polypeptide Homo sapiens mucin-type O-linked glycosylation of serine or threonine residues UDP + N-acetyl-D-galactosaminyl-polypeptide
-
 ?

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
 isoforms ppGalNAcT-2 (hT2) and ppGalNAcT-10 (hT10)
-

Source Tissue

Source Tissue Comment Organism Textmining
HEK-293 cell ppGalNAcT-10 amplified from isolated total RNA Homo sapiens
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
MUC5AC + UDP-GalNAc hT2CD, hT2: glycosylation site selection driven by and substrate binding through its catalytic domain, hT10: not catalysed by hT10 because hT10 does not recognize naked peptides Homo sapiens GalNAc-Thr9-MUC5AC + UDP preferred site for glycosylation by hT2 and hT2CD compared to Thr-3, Thr-10 and Thr-13 ?
MUC5AC glycopeptides + UDP-GalNAc hT2: site preference of glycosylation depends on location of pre-existing GalNAc and is mediated by its lectin domain which directs glycosylation 10 residues N- or C-terminal to an extant GalNAc residue and aids site selection only when potential site is N-terminal to an extant GalNAc residue not C-terminal, Thr-9 is no preferred site of glycosylation, hT10: must recognize existing GalNAc residues via its catalytic domain which directs selection to the glycosylation site immediately N-terminal to the present GalNAc, Homo sapiens GalNAc-MUC5AC + UDP
-
 ?
MUC5AC-13 + UDP-GalNAc hT10, hT10CD, hT10CD-hT2LD Homo sapiens GalNAc-Thr12-MUC5AC-13 + UDP
-
 ?
MUC5AC-13 + UDP-GalNAc hT2: substrate binding through its catalytic domain, hT10: transfer of only single GalNAc residues Homo sapiens GalNAc-Thr3-MUC5AC-13 + UDP predominant product of hT2 and hT10 ?
MUC5AC-13 + UDP-GalNAc hT2, hT2CD: no initial burst phase during catalysis Homo sapiens GalNAc-Thr9-MUC5AC-13 + UDP not preferred by hT2, preferred by hT2CD ?
MUC5AC-3 + UDP-GalNAc hT2: no initial burst phase during catalysis, substrate binding through its catalytic domain, hT10: transfer of only single GalNAc residues Homo sapiens GalNAc-Thr13-MUC5AC-3 + UDP hT2: preferred site for glycosylation, mediated by enhanced product release through lectin domain, hT10: predominant product ?
MUC5AC-3 + UDP-GalNAc hT10CD, hT10CD-hT2LD, hT10: catalysis faster than for MUCAC-9 Homo sapiens GalNAc-Thr2-MUC5AC + UDP
-
 ?
MUC5AC-3 + UDP-GalNAc hT2, hT2CD Homo sapiens GalNAc-Thr9-MUC5AC-3 + UDP not preferred by hT2, hT2CD: preferred site for glycosylation, lack of lectin domain in hT2CD results in rate-limiting product release ?
MUC5AC-3,13 + UDP-GalNAc hT2, not catalysed by hT2CD and hT2CD-hT10LD Homo sapiens GalNAc-Ser-5-MUC5AC-3,13 + UDP preferred site by hT2 ?
MUC5AC-3,13 + UDP-GalNAc hT10, hT10CD, hT10CD-hT2LD Homo sapiens GalNAc-Thr2,12-MUC5AC-3,13 + UDP
-
 ?
MUC5AC-9 + UDP-GalNAc not catalysed by hT10CD and hT10CD-hT2LD, hT10: catalysis slower than for MUC5AC-3 Homo sapiens GalNAc-Thr2-MUC5AC-9 + UDP preferred site of glycosylation by hT10 ?
UDP-N-acetyl-D-galactosamine + polypeptide mucin-type O-linked glycosylation of serine or threonine residues Homo sapiens UDP + N-acetyl-D-galactosaminyl-polypeptide
-
 ?

Synonyms

Synonyms Comment Organism
ppGalNAcT or hT
-
 Homo sapiens
UDP-GalNAc:polypeptide alpha-N-acetylgalactosaminyltransferase
-
 Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
additional information
-
 additional information hT2 catalysed glycosylation of MUC5AC and MUC5AC-13 follows a biphasic time course: fast formation of enzyme-product (initial burst phase: 1 molecule product per molecule hT2 enzyme), slow and rate-limiting product release Homo sapiens
additional information
-
 additional information hT2 catalysed glycosylation of MUC5AC-3 follows a linear time course: rate limiting catalysis or substrate binding Homo sapiens
additional information
-
 additional information steady-state rates of hT2 catalysis are less than 1 reaction per second at room temperature Homo sapiens
0.04
-
 MUC5AC at site Thr-9, hT2, at pre-steady state Homo sapiens
0.04
-
 MUC5AC-3 at site Thr-9, hT2CD, at pre-steady state Homo sapiens
0.05
-
 MUC5AC at site Thr-9, hT2CD Homo sapiens
0.052
-
 MUC5AC at site Thr-9, hT2 Homo sapiens
0.053
-
 MUC5AC at site Thr-9, hT2CD, at pre-steady state Homo sapiens
0.058
-
 MUC5AC-3 at site Thr-9, hT2CD Homo sapiens
0.067
-
 MUC5AC-13 at site Thr-9, hT2CD Homo sapiens
0.075
-
 MUC5AC-13 at site Thr-9, hT2CD, at pre-steady state Homo sapiens
0.103
-
 MUC5AC-13 at site Thr-3, hT2 Homo sapiens
0.108
-
 MUC5AC-13 at site Thr-3, hT2, at pre-steady state Homo sapiens
0.123
-
 MUC5AC at site Thr-9, hT2, single turnover event Homo sapiens
0.127
-
 MUC5AC-9 at site Thr-2, hT10 Homo sapiens
0.137
-
 MUC5AC-3 at site Thr-9, hT2CD, single turnover event Homo sapiens
0.15
-
 MUC5AC-3 at site Thr-2, hT10CD-hT2LD Homo sapiens
0.17
-
 MUC5AC-13 at site Thr-3, hT2, single turnover event Homo sapiens
0.193
-
 MUC5AC at site Thr-9, hT2CD, single turnover event Homo sapiens
0.218
-
 MUC5AC-3 at site Thr-13, hT2 Homo sapiens
0.22
-
 MUC5AC-3 at site Thr-13, hT2, at pre-steady state Homo sapiens
0.24
-
 MUC5AC-3 at site Thr-2, hT10CD Homo sapiens
0.29
-
 MUC5AC-13 at site Thr-12, hT10CD-hT2LD Homo sapiens
0.31
-
 MUC5AC-13 at site Thr-12, hT10CD Homo sapiens
0.33
-
 MUC5AC-3 at site Thr-2, hT10 Homo sapiens
0.4
-
 MUC5AC-13 at site Thr-12, hT10 Homo sapiens
0.52
-
 MUC5AC-3,13 at site Thr-2 and Thr-12, hT10CD-hT2LD Homo sapiens
0.76
-
 MUC5AC-3,13 at sites Thr-2 and Thr-12, hT10CD Homo sapiens
1.4
-
 MUC5AC-3,13 at site Thr-2 and Thr-12, hT10 Homo sapiens