Cargando…
Thermodynamic Switch in Binding of Adhesion/Growth Regulatory Human Galectin-3 to Tumor-Associated TF Antigen (CD176) and MUC1 Glycopeptides
[Image: see text] A shift to short-chain glycans is an observed change in mucin-type O-glycosylation in premalignant and malignant epithelia. Given the evidence that human galectin-3 can interact with mucins and also weakly with free tumor-associated Thomsen-Friedenreich (TF) antigen (CD176), the st...
Autores principales: | , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American
Chemical Society
2015
|
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4520625/ https://www.ncbi.nlm.nih.gov/pubmed/26129647 http://dx.doi.org/10.1021/acs.biochem.5b00555 |
_version_ | 1782383693637091328 |
---|---|
author | Rodriguez, Maria C. Yegorova, Svetlana Pitteloud, Jean-Philippe Chavaroche, Anais E. André, Sabine Ardá, Ana Minond, Dimitriy Jiménez-Barbero, Jesús Gabius, Hans-Joachim Cudic, Mare |
author_facet | Rodriguez, Maria C. Yegorova, Svetlana Pitteloud, Jean-Philippe Chavaroche, Anais E. André, Sabine Ardá, Ana Minond, Dimitriy Jiménez-Barbero, Jesús Gabius, Hans-Joachim Cudic, Mare |
author_sort | Rodriguez, Maria C. |
collection | PubMed |
description | [Image: see text] A shift to short-chain glycans is an observed change in mucin-type O-glycosylation in premalignant and malignant epithelia. Given the evidence that human galectin-3 can interact with mucins and also weakly with free tumor-associated Thomsen-Friedenreich (TF) antigen (CD176), the study of its interaction with MUC1 (glyco)peptides is of biomedical relevance. Glycosylated MUC1 fragments that carry the TF antigen attached through either Thr or Ser side chains were synthesized using standard Fmoc-based automated solid-phase peptide chemistry. The dissociation constants (K(d)) for interaction of galectin-3 and the glycosylated MUC1 fragments measured by isothermal titration calorimetry decreased up to 10 times in comparison to that of the free TF disaccharide. No binding was observed for the nonglycosylated control version of the MUC1 peptide. The most notable feature of the binding of MUC1 glycopeptides to galectin-3 was a shift from a favorable enthalpy to an entropy-driven binding process. The comparatively diminished enthalpy contribution to the free energy (ΔG) was compensated by a considerable gain in the entropic term. (1)H–(15)N heteronuclear single-quantum coherence spectroscopy nuclear magnetic resonance data reveal contact at the canonical site mainly by the glycan moiety of the MUC1 glycopeptide. Ligand-dependent differences in binding affinities were also confirmed by a novel assay for screening of low-affinity glycan–lectin interactions based on AlphaScreen technology. Another key finding is that the glycosylated MUC1 peptides exhibited activity in a concentration-dependent manner in cell-based assays revealing selectivity among human galectins. Thus, the presentation of this tumor-associated carbohydrate ligand by the natural peptide scaffold enhances its affinity, highlighting the significance of model studies of human lectins with synthetic glycopeptides. |
format | Online Article Text |
id | pubmed-4520625 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | American
Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-45206252016-07-01 Thermodynamic Switch in Binding of Adhesion/Growth Regulatory Human Galectin-3 to Tumor-Associated TF Antigen (CD176) and MUC1 Glycopeptides Rodriguez, Maria C. Yegorova, Svetlana Pitteloud, Jean-Philippe Chavaroche, Anais E. André, Sabine Ardá, Ana Minond, Dimitriy Jiménez-Barbero, Jesús Gabius, Hans-Joachim Cudic, Mare Biochemistry [Image: see text] A shift to short-chain glycans is an observed change in mucin-type O-glycosylation in premalignant and malignant epithelia. Given the evidence that human galectin-3 can interact with mucins and also weakly with free tumor-associated Thomsen-Friedenreich (TF) antigen (CD176), the study of its interaction with MUC1 (glyco)peptides is of biomedical relevance. Glycosylated MUC1 fragments that carry the TF antigen attached through either Thr or Ser side chains were synthesized using standard Fmoc-based automated solid-phase peptide chemistry. The dissociation constants (K(d)) for interaction of galectin-3 and the glycosylated MUC1 fragments measured by isothermal titration calorimetry decreased up to 10 times in comparison to that of the free TF disaccharide. No binding was observed for the nonglycosylated control version of the MUC1 peptide. The most notable feature of the binding of MUC1 glycopeptides to galectin-3 was a shift from a favorable enthalpy to an entropy-driven binding process. The comparatively diminished enthalpy contribution to the free energy (ΔG) was compensated by a considerable gain in the entropic term. (1)H–(15)N heteronuclear single-quantum coherence spectroscopy nuclear magnetic resonance data reveal contact at the canonical site mainly by the glycan moiety of the MUC1 glycopeptide. Ligand-dependent differences in binding affinities were also confirmed by a novel assay for screening of low-affinity glycan–lectin interactions based on AlphaScreen technology. Another key finding is that the glycosylated MUC1 peptides exhibited activity in a concentration-dependent manner in cell-based assays revealing selectivity among human galectins. Thus, the presentation of this tumor-associated carbohydrate ligand by the natural peptide scaffold enhances its affinity, highlighting the significance of model studies of human lectins with synthetic glycopeptides. American Chemical Society 2015-07-01 2015-07-28 /pmc/articles/PMC4520625/ /pubmed/26129647 http://dx.doi.org/10.1021/acs.biochem.5b00555 Text en Copyright © 2015 American Chemical Society This is an open access article published under an ACS AuthorChoice License (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html) , which permits copying and redistribution of the article or any adaptations for non-commercial purposes. |
spellingShingle | Rodriguez, Maria C. Yegorova, Svetlana Pitteloud, Jean-Philippe Chavaroche, Anais E. André, Sabine Ardá, Ana Minond, Dimitriy Jiménez-Barbero, Jesús Gabius, Hans-Joachim Cudic, Mare Thermodynamic Switch in Binding of Adhesion/Growth Regulatory Human Galectin-3 to Tumor-Associated TF Antigen (CD176) and MUC1 Glycopeptides |
title | Thermodynamic Switch in Binding of Adhesion/Growth Regulatory Human Galectin-3 to Tumor-Associated TF Antigen (CD176) and MUC1 Glycopeptides |
title_full | Thermodynamic Switch in Binding of Adhesion/Growth Regulatory Human Galectin-3 to Tumor-Associated TF Antigen (CD176) and MUC1 Glycopeptides |
title_fullStr | Thermodynamic Switch in Binding of Adhesion/Growth Regulatory Human Galectin-3 to Tumor-Associated TF Antigen (CD176) and MUC1 Glycopeptides |
title_full_unstemmed | Thermodynamic Switch in Binding of Adhesion/Growth Regulatory Human Galectin-3 to Tumor-Associated TF Antigen (CD176) and MUC1 Glycopeptides |
title_short | Thermodynamic Switch in Binding of Adhesion/Growth Regulatory Human Galectin-3 to Tumor-Associated TF Antigen (CD176) and MUC1 Glycopeptides |
title_sort | thermodynamic switch in binding of adhesion/growth regulatory human galectin-3 to tumor-associated tf antigen (cd176) and muc1 glycopeptides |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4520625/ https://www.ncbi.nlm.nih.gov/pubmed/26129647 http://dx.doi.org/10.1021/acs.biochem.5b00555 |
work_keys_str_mv | AT rodriguezmariac thermodynamicswitchinbindingofadhesiongrowthregulatoryhumangalectin3totumorassociatedtfantigencd176andmuc1glycopeptides AT yegorovasvetlana thermodynamicswitchinbindingofadhesiongrowthregulatoryhumangalectin3totumorassociatedtfantigencd176andmuc1glycopeptides AT pitteloudjeanphilippe thermodynamicswitchinbindingofadhesiongrowthregulatoryhumangalectin3totumorassociatedtfantigencd176andmuc1glycopeptides AT chavarocheanaise thermodynamicswitchinbindingofadhesiongrowthregulatoryhumangalectin3totumorassociatedtfantigencd176andmuc1glycopeptides AT andresabine thermodynamicswitchinbindingofadhesiongrowthregulatoryhumangalectin3totumorassociatedtfantigencd176andmuc1glycopeptides AT ardaana thermodynamicswitchinbindingofadhesiongrowthregulatoryhumangalectin3totumorassociatedtfantigencd176andmuc1glycopeptides AT minonddimitriy thermodynamicswitchinbindingofadhesiongrowthregulatoryhumangalectin3totumorassociatedtfantigencd176andmuc1glycopeptides AT jimenezbarberojesus thermodynamicswitchinbindingofadhesiongrowthregulatoryhumangalectin3totumorassociatedtfantigencd176andmuc1glycopeptides AT gabiushansjoachim thermodynamicswitchinbindingofadhesiongrowthregulatoryhumangalectin3totumorassociatedtfantigencd176andmuc1glycopeptides AT cudicmare thermodynamicswitchinbindingofadhesiongrowthregulatoryhumangalectin3totumorassociatedtfantigencd176andmuc1glycopeptides |