Glycopeptide-preferring polypeptide GalNAc transferase 10 (ppGalNAc T10), involved in mucin-type O-glycosylation, has a unique GalNAc-O-Ser/Thr-binding site in its catalytic domain not found in ppGalNAc T1 or T2

Cynthia L. Perrine, Anjali Ganguli, Peng Wu, Carolyn R. Bertozzi, Timothy A. Fritz, Jayalakshmi Raman, Lawrence A. Tabak, Thomas A. Gerken

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Mucin-type O-glycosylation is initiated by a large family of UDP-GalNAc:polypeptide α-N-acetylgalactosaminyltransferases (ppGalNAc Ts) that transfer GalNAc from UDP-GalNAc to the Ser and Thr residues of polypeptide acceptors. Some members of the family prefer previously glycosylated peptides (ppGalNAc T7 and T10), whereas others are inhibited by neighboring glycosylation (ppGalNAc T1 and T2). Characterizing their peptide and glycopeptide substrate specificity is critical for understanding the biological role and significance of each isoform. Utilizing a series of random peptide and glycopeptide substrates, we have obtained the peptide and glycopeptide specificities of ppGalNAc T10 for comparison with ppGalNAc T1 and T2. For the glycopeptide substrates, ppGalNAc T10 exhibited a single large preference for Ser/Thr-O-GalNAc at the b1 (C-terminal) position relative to the Ser or Thr acceptor site. ppGalNAc T1 and T2 revealed no significant enhancements suggesting Ser/Thr-O-GalNAc was inhibitory at most positions for these isoforms. Against random peptide substrates, ppGalNAc T10 revealed no significant hydrophobic or hydrophilic residue enhancements, in contrast to what has been reported previously for ppGal-NAc T1 and T2. Our results reveal that these transferases have unique peptide and glycopeptide preferences demonstrating their substrate diversity and their likely roles ranging from initiating transferases to filling-in transferases.

Original languageEnglish (US)
Pages (from-to)20387-20397
Number of pages11
JournalJournal of Biological Chemistry
Volume284
Issue number30
DOIs
StatePublished - Jul 24 2009

Fingerprint

Glycosylation
Glycopeptides
Mucins
Catalytic Domain
Binding Sites
Peptides
Transferases
Substrates
Protein Isoforms
polypeptide N-acetylgalactosaminyltransferase
Uridine Diphosphate
Substrate Specificity

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Glycopeptide-preferring polypeptide GalNAc transferase 10 (ppGalNAc T10), involved in mucin-type O-glycosylation, has a unique GalNAc-O-Ser/Thr-binding site in its catalytic domain not found in ppGalNAc T1 or T2. / Perrine, Cynthia L.; Ganguli, Anjali; Wu, Peng; Bertozzi, Carolyn R.; Fritz, Timothy A.; Raman, Jayalakshmi; Tabak, Lawrence A.; Gerken, Thomas A.

In: Journal of Biological Chemistry, Vol. 284, No. 30, 24.07.2009, p. 20387-20397.

Research output: Contribution to journalArticle

Perrine, Cynthia L. ; Ganguli, Anjali ; Wu, Peng ; Bertozzi, Carolyn R. ; Fritz, Timothy A. ; Raman, Jayalakshmi ; Tabak, Lawrence A. ; Gerken, Thomas A. / Glycopeptide-preferring polypeptide GalNAc transferase 10 (ppGalNAc T10), involved in mucin-type O-glycosylation, has a unique GalNAc-O-Ser/Thr-binding site in its catalytic domain not found in ppGalNAc T1 or T2. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 30. pp. 20387-20397.
@article{7718ff3b1aa34b08a1f483f40bf9e2b6,
title = "Glycopeptide-preferring polypeptide GalNAc transferase 10 (ppGalNAc T10), involved in mucin-type O-glycosylation, has a unique GalNAc-O-Ser/Thr-binding site in its catalytic domain not found in ppGalNAc T1 or T2",
abstract = "Mucin-type O-glycosylation is initiated by a large family of UDP-GalNAc:polypeptide α-N-acetylgalactosaminyltransferases (ppGalNAc Ts) that transfer GalNAc from UDP-GalNAc to the Ser and Thr residues of polypeptide acceptors. Some members of the family prefer previously glycosylated peptides (ppGalNAc T7 and T10), whereas others are inhibited by neighboring glycosylation (ppGalNAc T1 and T2). Characterizing their peptide and glycopeptide substrate specificity is critical for understanding the biological role and significance of each isoform. Utilizing a series of random peptide and glycopeptide substrates, we have obtained the peptide and glycopeptide specificities of ppGalNAc T10 for comparison with ppGalNAc T1 and T2. For the glycopeptide substrates, ppGalNAc T10 exhibited a single large preference for Ser/Thr-O-GalNAc at the b1 (C-terminal) position relative to the Ser or Thr acceptor site. ppGalNAc T1 and T2 revealed no significant enhancements suggesting Ser/Thr-O-GalNAc was inhibitory at most positions for these isoforms. Against random peptide substrates, ppGalNAc T10 revealed no significant hydrophobic or hydrophilic residue enhancements, in contrast to what has been reported previously for ppGal-NAc T1 and T2. Our results reveal that these transferases have unique peptide and glycopeptide preferences demonstrating their substrate diversity and their likely roles ranging from initiating transferases to filling-in transferases.",
author = "Perrine, {Cynthia L.} and Anjali Ganguli and Peng Wu and Bertozzi, {Carolyn R.} and Fritz, {Timothy A.} and Jayalakshmi Raman and Tabak, {Lawrence A.} and Gerken, {Thomas A.}",
year = "2009",
month = "7",
day = "24",
doi = "10.1074/jbc.M109.017236",
language = "English (US)",
volume = "284",
pages = "20387--20397",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "30",

}

TY - JOUR

T1 - Glycopeptide-preferring polypeptide GalNAc transferase 10 (ppGalNAc T10), involved in mucin-type O-glycosylation, has a unique GalNAc-O-Ser/Thr-binding site in its catalytic domain not found in ppGalNAc T1 or T2

AU - Perrine, Cynthia L.

AU - Ganguli, Anjali

AU - Wu, Peng

AU - Bertozzi, Carolyn R.

AU - Fritz, Timothy A.

AU - Raman, Jayalakshmi

AU - Tabak, Lawrence A.

AU - Gerken, Thomas A.

PY - 2009/7/24

Y1 - 2009/7/24

N2 - Mucin-type O-glycosylation is initiated by a large family of UDP-GalNAc:polypeptide α-N-acetylgalactosaminyltransferases (ppGalNAc Ts) that transfer GalNAc from UDP-GalNAc to the Ser and Thr residues of polypeptide acceptors. Some members of the family prefer previously glycosylated peptides (ppGalNAc T7 and T10), whereas others are inhibited by neighboring glycosylation (ppGalNAc T1 and T2). Characterizing their peptide and glycopeptide substrate specificity is critical for understanding the biological role and significance of each isoform. Utilizing a series of random peptide and glycopeptide substrates, we have obtained the peptide and glycopeptide specificities of ppGalNAc T10 for comparison with ppGalNAc T1 and T2. For the glycopeptide substrates, ppGalNAc T10 exhibited a single large preference for Ser/Thr-O-GalNAc at the b1 (C-terminal) position relative to the Ser or Thr acceptor site. ppGalNAc T1 and T2 revealed no significant enhancements suggesting Ser/Thr-O-GalNAc was inhibitory at most positions for these isoforms. Against random peptide substrates, ppGalNAc T10 revealed no significant hydrophobic or hydrophilic residue enhancements, in contrast to what has been reported previously for ppGal-NAc T1 and T2. Our results reveal that these transferases have unique peptide and glycopeptide preferences demonstrating their substrate diversity and their likely roles ranging from initiating transferases to filling-in transferases.

AB - Mucin-type O-glycosylation is initiated by a large family of UDP-GalNAc:polypeptide α-N-acetylgalactosaminyltransferases (ppGalNAc Ts) that transfer GalNAc from UDP-GalNAc to the Ser and Thr residues of polypeptide acceptors. Some members of the family prefer previously glycosylated peptides (ppGalNAc T7 and T10), whereas others are inhibited by neighboring glycosylation (ppGalNAc T1 and T2). Characterizing their peptide and glycopeptide substrate specificity is critical for understanding the biological role and significance of each isoform. Utilizing a series of random peptide and glycopeptide substrates, we have obtained the peptide and glycopeptide specificities of ppGalNAc T10 for comparison with ppGalNAc T1 and T2. For the glycopeptide substrates, ppGalNAc T10 exhibited a single large preference for Ser/Thr-O-GalNAc at the b1 (C-terminal) position relative to the Ser or Thr acceptor site. ppGalNAc T1 and T2 revealed no significant enhancements suggesting Ser/Thr-O-GalNAc was inhibitory at most positions for these isoforms. Against random peptide substrates, ppGalNAc T10 revealed no significant hydrophobic or hydrophilic residue enhancements, in contrast to what has been reported previously for ppGal-NAc T1 and T2. Our results reveal that these transferases have unique peptide and glycopeptide preferences demonstrating their substrate diversity and their likely roles ranging from initiating transferases to filling-in transferases.

UR - http://www.scopus.com/inward/record.url?scp=67749133871&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=67749133871&partnerID=8YFLogxK

U2 - 10.1074/jbc.M109.017236

DO - 10.1074/jbc.M109.017236

M3 - Article

C2 - 19460755

AN - SCOPUS:67749133871

VL - 284

SP - 20387

EP - 20397

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 30

ER -