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PMID: 15486088 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The beginnings of mucin biosynthesis: the crystal structure of UDP-GalNAc:polypeptide alpha-N-acetylgalactosaminyltransferase-T1.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 101 ·No. 43 ·2004-10-26 ·Pages 15307-12

Fritz TA, Hurley JH, Trinh LB, Shiloach J, Tabak LA

Abstract

UDP-GalNAc:polypeptide alpha-N-acetylgalactosaminyltransferases (ppGaNTases) initiate the formation of mucin-type, O-linked glycans by catalyzing the transfer of alpha-N-acetylgalactosamine from UDP-GalNAc to Ser or Thr residues of core proteins to form the Tn antigen (GalNAc-alpha-1-O-Ser/Thr). ppGaNTases are unique among glycosyltransferases in containing a C-terminal lectin domain. We present the x-ray crystal structure of a ppGaNTase, murine ppGaNTase-T1, and show that it folds to form distinct catalytic and lectin domains. The association of the two domains forms a large cleft in the surface of the enzyme that contains a Mn2+ ion complexed by invariant D209 and H211 of the "DXH" motif and by invariant H344. Each of the three potential lectin domain carbohydrate-binding sites (alpha, beta, and gamma) is located on the active-site face of the enzyme, suggesting a mechanism by which the transferase may accommodate multiple conformations of glycosylated acceptor substrates. A model of a mucin 1 glycopeptide substrate bound to the enzyme shows that the spatial separation between the lectin alpha site and a modeled active site UDP-GalNAc is consistent with the in vitro pattern of glycosylation observed for this peptide catalyzed by ppGaNTase-T1. The structure also provides a template for the larger ppGaNTase family, and homology models of several ppGaNTase isoforms predict dramatically different surface chemistries consistent with isoform-selective acceptor substrate recognition.

MeSH Terms
Animals Binding Sites Catalytic Domain Crystallography, X-Ray Lectins/metabolism Mice Models, Molecular Mucins/biosynthesis N-Acetylgalactosaminyltransferases/chemistry,genetics,metabolism Protein Conformation
Chemicals
Lectins Mucins N-Acetylgalactosaminyltransferases polypeptide N-acetylgalactosaminyltransferase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Fritz Timothy A
Section on Biological Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Department of Health and Human Services, Bethesda, MD 20892, USA.
Hurley James H
Trinh Loc-Ba
Shiloach Joseph
Tabak Lawrence A
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0405657101
Published
2004-10-26
Epub
2004-00-14
Pages
15307-12
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC524453
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.0405657101 PMC Full Text View journal details
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