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Literature summary for 2.4.1.41 extracted from

  • Choi, J.; Wagner, L.J.S.; Timmermans, S.B.P.E.; Malaker, S.A.; Schumann, B.; Gray, M.A.; Debets, M.F.; Takashima, M.; Gehring, J.; Bertozzi, C.R.
    Engineering orthogonal polypeptide GalNAc-transferase and UDP-sugar pairs (2019), J. Am. Chem. Soc., 141, 13442-13453 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.4.1.41

Application

Application Comment Organism
analysis development of a bump-hole chemical reporter system for studying GalNAc-T activity in vitro. GalNAc-T2 ire rationally engineered to contain an enlarged active site (hole) and probed with a collection of 20 (bumped) uridine diphosphate N-acetylgalactosamine analogs to identify enzyme-substrate pairs that retain peptide specificities but are otherwise completely orthogonal to native enzyme-substrate pairs. The approach is applicable to multiple GalNAc-T isoenzymes, including GalNAc-T1 and -T2 that prefer nonglycosylated peptide substrates and GalNAcT-10 that prefers a preglycosylated peptide substrate Homo sapiens

Protein Variants

Protein Variants Comment Organism
F361A minimal residual activity Homo sapiens
F361S minimal residual activity Homo sapiens
I253A reduced activity with substrate UDP-GalNAc Homo sapiens
I253A/L310A UDP-GalNAc is a poor substrate, some UDP-GalNAc analogs with longer or branched N-acyl chains are better substrates for mutant I253A/L310A than for wild-type Homo sapiens
L310A reduced activity with substrate UDP-GalNAc Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0026
-
 UDP-N-hex-5-ynoyl-alpha-D-galactosamine mutant 253A/L310A, pH 7.4, 37°C Homo sapiens
0.03
-
 UDP-N-acetyl-alpha-D-galactosamine wild-type, pH 7.4, 37°C Homo sapiens
0.043
-
 UDP-N-((2S)-2-azidopropanoyl)-alpha-D-galactosamine mutant 253A/L310A, pH 7.4, 37°C Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens Q10471 isoform GalNAc-T2
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
UDP-N-((2S)-2-azidopropanoyl)-alpha-D-galactosamine + peptide EA2-biotin i.e. PTTDSTTPAPTTKK-biotin Homo sapiens UDP + N-((2S)-2-azidopropanoyl)-alpha-D-galactosaminyl-peptide EA2-biotin
-
-
UDP-N-acetyl-alpha-D-galactosamine + peptide EA2-biotin i.e. PTTDSTTPAPTTKK-biotin Homo sapiens UDP + N-acetyl-alpha-D-galactosaminyl-peptide EA2-biotin
-
-
UDP-N-hex-5-ynoyl-alpha-D-galactosamine + peptide EA2-biotin i.e. PTTDSTTPAPTTKK-biotin Homo sapiens UDP + N-hex-5-ynoyl-alpha-D-galactosaminyl-peptide EA2-biotin
-
-

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.158
-
 UDP-N-hex-5-ynoyl-alpha-D-galactosamine mutant 253A/L310A, pH 7.4, 37°C Homo sapiens
0.566
-
 UDP-N-((2S)-2-azidopropanoyl)-alpha-D-galactosamine mutant 253A/L310A, pH 7.4, 37°C Homo sapiens
0.813
-
 UDP-N-acetyl-alpha-D-galactosamine wild-type, pH 7.4, 37°C Homo sapiens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
13
-
 UDP-N-((2S)-2-azidopropanoyl)-alpha-D-galactosamine mutant 253A/L310A, pH 7.4, 37°C Homo sapiens
28
-
 UDP-N-acetyl-alpha-D-galactosamine wild-type, pH 7.4, 37°C Homo sapiens
61
-
 UDP-N-hex-5-ynoyl-alpha-D-galactosamine mutant 253A/L310A, pH 7.4, 37°C Homo sapiens