The structure of the colorectal cancer-associated enzyme GalNAc-T12 reveals how nonconserved residues dictate its function
Abstract
Polypeptide N-acetylgalactosaminyl transferases (GalNAc-Ts) initiate mucin type O-glycosylation by catalyzing the transfer of N-acetylgalactosamine (GalNAc) to Ser or Thr on a protein substrate. Inactive and partially active variants of the isoenzyme GalNAc-T12 are present in subsets of patients with colorectal cancer, and several of these variants alter nonconserved residues with unknown functions. While previous biochemical studies have demonstrated that GalNAc-T12 selects for peptide and glycopeptide substrates through unique interactions with its catalytic and lectin domains, the molecular basis for this distinct substrate selectivity remains elusive. Here we examine the molecular basis of the activity and substrate selectivity of GalNAc-T12. The X-ray crystal structure of GalNAc-T12 in complex with a di-glycosylated peptide substrate reveals how a nonconserved GalNAc binding pocket in the GalNAc-T12 catalytic domain dictates its unique substrate selectivity. In addition, the structure provides insight into how colorectal cancer mutations disrupt the activity of GalNAc-T12 and illustrates how the rules dictating GalNAc-T12 function are distinct from those for other GalNAc-Ts.
- Authors:
-
[3];
[1]
- National Inst. of Health (NIH), Bethesda, MD (United States). National Inst. of Dental and Craniofacial Research
- Case Western Reserve Univ., Cleveland, OH (United States)
- Johns Hopkins Univ., Baltimore, MD (United States)
- Publication Date:
- Research Org.:
- Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
- Sponsoring Org.:
- National Institutes of Health (NIH)
- OSTI Identifier:
- 1576004
- Grant/Contract Number:
- 1-ZIADE000739-05; R01 GM113534
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Proceedings of the National Academy of Sciences of the United States of America
- Additional Journal Information:
- Journal Volume: 116; Journal Issue: 41; Journal ID: ISSN 0027-8424
- Publisher:
- National Academy of Sciences
- Country of Publication:
- United States
- Language:
- ENGLISH
- Subject:
- 60 APPLIED LIFE SCIENCES; GalNAc-Ts; mucin-type O-glycosylation; colorectal cancer; substrate selectivity; enzyme catalysis
Citation Formats
Fernandez, Amy J., Daniel, Earnest James Paul, Mahajan, Sai Pooja, Gray, Jeffrey J., Gerken, Thomas A., Tabak, Lawrence A., and Samara, Nadine L. The structure of the colorectal cancer-associated enzyme GalNAc-T12 reveals how nonconserved residues dictate its function. United States: N. p., 2019.
Web. doi:10.1073/pnas.1902211116.
Fernandez, Amy J., Daniel, Earnest James Paul, Mahajan, Sai Pooja, Gray, Jeffrey J., Gerken, Thomas A., Tabak, Lawrence A., & Samara, Nadine L. The structure of the colorectal cancer-associated enzyme GalNAc-T12 reveals how nonconserved residues dictate its function. United States. https://doi.org/10.1073/pnas.1902211116
Fernandez, Amy J., Daniel, Earnest James Paul, Mahajan, Sai Pooja, Gray, Jeffrey J., Gerken, Thomas A., Tabak, Lawrence A., and Samara, Nadine L. Mon .
"The structure of the colorectal cancer-associated enzyme GalNAc-T12 reveals how nonconserved residues dictate its function". United States. https://doi.org/10.1073/pnas.1902211116. https://www.osti.gov/servlets/purl/1576004.
@article{osti_1576004,
title = {The structure of the colorectal cancer-associated enzyme GalNAc-T12 reveals how nonconserved residues dictate its function},
author = {Fernandez, Amy J. and Daniel, Earnest James Paul and Mahajan, Sai Pooja and Gray, Jeffrey J. and Gerken, Thomas A. and Tabak, Lawrence A. and Samara, Nadine L.},
abstractNote = {Polypeptide N-acetylgalactosaminyl transferases (GalNAc-Ts) initiate mucin type O-glycosylation by catalyzing the transfer of N-acetylgalactosamine (GalNAc) to Ser or Thr on a protein substrate. Inactive and partially active variants of the isoenzyme GalNAc-T12 are present in subsets of patients with colorectal cancer, and several of these variants alter nonconserved residues with unknown functions. While previous biochemical studies have demonstrated that GalNAc-T12 selects for peptide and glycopeptide substrates through unique interactions with its catalytic and lectin domains, the molecular basis for this distinct substrate selectivity remains elusive. Here we examine the molecular basis of the activity and substrate selectivity of GalNAc-T12. The X-ray crystal structure of GalNAc-T12 in complex with a di-glycosylated peptide substrate reveals how a nonconserved GalNAc binding pocket in the GalNAc-T12 catalytic domain dictates its unique substrate selectivity. In addition, the structure provides insight into how colorectal cancer mutations disrupt the activity of GalNAc-T12 and illustrates how the rules dictating GalNAc-T12 function are distinct from those for other GalNAc-Ts.},
doi = {10.1073/pnas.1902211116},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 41,
volume = 116,
place = {United States},
year = {2019},
month = {9}
}
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