Delineating binding modes of Gal/GalNAc and structural elements of the molecular recognition of tumor-associated mucin glycopeptides by the human macrophage galactose-type lectin

Filipa Marcelo, Fayna Garcia-Martin, Takahiko Matsushita, João Sardinha, Helena Coelho, Anneloes Oude-Vrielink, Christiane Koller, Sabine André, Eurico J. Cabrita, Hans Joachim Gabius, Shin Ichiro Nishimura, Jesffls Jiménez-Barbero, F. Javier Cañada

Research output: Contribution to journalArticle

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Abstract

The human macrophage galactose-type lectin (MGL) is a key physiological receptor for the carcinoma-associated Tn antigen (GalNAc-α-1-O-Ser/Thr) in mucins. NMR and modeling-based data on the molecular recognition features of synthetic Tn-bearing glycopeptides by MGL are presented. Cognate epitopes on the sugar and matching key amino acids involved in the interaction were identified by saturation transfer difference (STD) NMR spectroscopy. Only the amino acids close to the glycosylation site in the peptides are involved in lectin contact. Moreover, control experi-ments with non-glycosylated MUC1 peptides unequivocally showed that the sugar residue is essential for MGL binding, as is Ca2+. NMR data were complemented with molecular dynamics simulations and Corcema-ST to establish a 3D view on the molecular recognition process between Gal, GalNAc, and the Tn-presenting glycopeptides and MGL. Gal and GalNAc have a dual binding mode with opposite trend of the main interaction pattern and the differences in affinity can be explained by additional hydrogen bonds and CH-p contacts involving exclusively the NHAc moiety.

Original languageEnglish
Pages (from-to)16147-16155
Number of pages9
JournalChemistry - A European Journal
Volume20
Issue number49
DOIs
Publication statusPublished - 12 Jan 2014

Fingerprint

Molecular recognition
Glycopeptides
Macrophages
Mucins
Lectins
Tumors
Galactose
Sugars
Bearings (structural)
Nuclear magnetic resonance
Galectins
Glycosylation
Amino Acids
Peptides
Nuclear magnetic resonance spectroscopy
Molecular dynamics
Epitopes
Hydrogen bonds
Computer simulation
human MGL lectin

Keywords

  • Binding studies
  • Macrophage galactose-type lectin
  • Molecular modeling
  • Molecular recognition
  • Mucin glycopeptides

Cite this

Marcelo, Filipa ; Garcia-Martin, Fayna ; Matsushita, Takahiko ; Sardinha, João ; Coelho, Helena ; Oude-Vrielink, Anneloes ; Koller, Christiane ; André, Sabine ; Cabrita, Eurico J. ; Gabius, Hans Joachim ; Nishimura, Shin Ichiro ; Jiménez-Barbero, Jesffls ; Cañada, F. Javier. / Delineating binding modes of Gal/GalNAc and structural elements of the molecular recognition of tumor-associated mucin glycopeptides by the human macrophage galactose-type lectin. In: Chemistry - A European Journal. 2014 ; Vol. 20, No. 49. pp. 16147-16155.
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abstract = "The human macrophage galactose-type lectin (MGL) is a key physiological receptor for the carcinoma-associated Tn antigen (GalNAc-α-1-O-Ser/Thr) in mucins. NMR and modeling-based data on the molecular recognition features of synthetic Tn-bearing glycopeptides by MGL are presented. Cognate epitopes on the sugar and matching key amino acids involved in the interaction were identified by saturation transfer difference (STD) NMR spectroscopy. Only the amino acids close to the glycosylation site in the peptides are involved in lectin contact. Moreover, control experi-ments with non-glycosylated MUC1 peptides unequivocally showed that the sugar residue is essential for MGL binding, as is Ca2+. NMR data were complemented with molecular dynamics simulations and Corcema-ST to establish a 3D view on the molecular recognition process between Gal, GalNAc, and the Tn-presenting glycopeptides and MGL. Gal and GalNAc have a dual binding mode with opposite trend of the main interaction pattern and the differences in affinity can be explained by additional hydrogen bonds and CH-p contacts involving exclusively the NHAc moiety.",
keywords = "Binding studies, Macrophage galactose-type lectin, Molecular modeling, Molecular recognition, Mucin glycopeptides",
author = "Filipa Marcelo and Fayna Garcia-Martin and Takahiko Matsushita and Jo{\~a}o Sardinha and Helena Coelho and Anneloes Oude-Vrielink and Christiane Koller and Sabine Andr{\'e} and Cabrita, {Eurico J.} and Gabius, {Hans Joachim} and Nishimura, {Shin Ichiro} and Jesffls Jim{\'e}nez-Barbero and Ca{\~n}ada, {F. Javier}",
note = "Sem PDF. The authors thank FCT Portugal for a post-doc research grant (SFRH/BPD/65462/2009) and for the financial support of EXPL/QEQ/MED/0799/2012 project, CQFB Strategic Project PEst-C/EQB/LA0006/2013 and MINECO, Spain, for the financial support of project CTQ2012-32025. The NMR spectrometer at FCT-UNL is part of the National NMR Network (RNRMN) and is funded by FCT-Portugal RECI/BBB-BQB/0230/2012. The generous financial support by the EC-funded Marie Curie Initial Training Network GLYCOPHARM (PITN-GA-2012-317297) and the GLYCOHIT program (grant agreement 260600) are also gratefully acknowledged. F.G.M. acknowledges the Japanese Special Coordination Fund for Promotion of Science and Technology for accelerated innovation of the fostering system for female scientists and JSPS Wakatte B KAKENHI Grant Number 24710242. We also thank S. Oka and A. Tokumitsu for ESI-HRMS measurements and T. Hirose for amino acid analysis at Instrumental Analysis Division, Equipment Management Center, Creative Research Institution, Hokkaido University. MUC1 (glyco)peptides syntheses of this work was supported by Project for Developing Innovation Systems of the Japanese Ministry of Education, Culture, Science and Technology and JSPS KAKENHI Grant Number 25220206.",
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Marcelo, F, Garcia-Martin, F, Matsushita, T, Sardinha, J, Coelho, H, Oude-Vrielink, A, Koller, C, André, S, Cabrita, EJ, Gabius, HJ, Nishimura, SI, Jiménez-Barbero, J & Cañada, FJ 2014, 'Delineating binding modes of Gal/GalNAc and structural elements of the molecular recognition of tumor-associated mucin glycopeptides by the human macrophage galactose-type lectin', Chemistry - A European Journal, vol. 20, no. 49, pp. 16147-16155. https://doi.org/10.1002/chem.201404566

Delineating binding modes of Gal/GalNAc and structural elements of the molecular recognition of tumor-associated mucin glycopeptides by the human macrophage galactose-type lectin. / Marcelo, Filipa; Garcia-Martin, Fayna; Matsushita, Takahiko; Sardinha, João; Coelho, Helena; Oude-Vrielink, Anneloes; Koller, Christiane; André, Sabine; Cabrita, Eurico J.; Gabius, Hans Joachim; Nishimura, Shin Ichiro; Jiménez-Barbero, Jesffls; Cañada, F. Javier.

In: Chemistry - A European Journal, Vol. 20, No. 49, 12.01.2014, p. 16147-16155.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Delineating binding modes of Gal/GalNAc and structural elements of the molecular recognition of tumor-associated mucin glycopeptides by the human macrophage galactose-type lectin

AU - Marcelo, Filipa

AU - Garcia-Martin, Fayna

AU - Matsushita, Takahiko

AU - Sardinha, João

AU - Coelho, Helena

AU - Oude-Vrielink, Anneloes

AU - Koller, Christiane

AU - André, Sabine

AU - Cabrita, Eurico J.

AU - Gabius, Hans Joachim

AU - Nishimura, Shin Ichiro

AU - Jiménez-Barbero, Jesffls

AU - Cañada, F. Javier

N1 - Sem PDF. The authors thank FCT Portugal for a post-doc research grant (SFRH/BPD/65462/2009) and for the financial support of EXPL/QEQ/MED/0799/2012 project, CQFB Strategic Project PEst-C/EQB/LA0006/2013 and MINECO, Spain, for the financial support of project CTQ2012-32025. The NMR spectrometer at FCT-UNL is part of the National NMR Network (RNRMN) and is funded by FCT-Portugal RECI/BBB-BQB/0230/2012. The generous financial support by the EC-funded Marie Curie Initial Training Network GLYCOPHARM (PITN-GA-2012-317297) and the GLYCOHIT program (grant agreement 260600) are also gratefully acknowledged. F.G.M. acknowledges the Japanese Special Coordination Fund for Promotion of Science and Technology for accelerated innovation of the fostering system for female scientists and JSPS Wakatte B KAKENHI Grant Number 24710242. We also thank S. Oka and A. Tokumitsu for ESI-HRMS measurements and T. Hirose for amino acid analysis at Instrumental Analysis Division, Equipment Management Center, Creative Research Institution, Hokkaido University. MUC1 (glyco)peptides syntheses of this work was supported by Project for Developing Innovation Systems of the Japanese Ministry of Education, Culture, Science and Technology and JSPS KAKENHI Grant Number 25220206.

PY - 2014/1/12

Y1 - 2014/1/12

N2 - The human macrophage galactose-type lectin (MGL) is a key physiological receptor for the carcinoma-associated Tn antigen (GalNAc-α-1-O-Ser/Thr) in mucins. NMR and modeling-based data on the molecular recognition features of synthetic Tn-bearing glycopeptides by MGL are presented. Cognate epitopes on the sugar and matching key amino acids involved in the interaction were identified by saturation transfer difference (STD) NMR spectroscopy. Only the amino acids close to the glycosylation site in the peptides are involved in lectin contact. Moreover, control experi-ments with non-glycosylated MUC1 peptides unequivocally showed that the sugar residue is essential for MGL binding, as is Ca2+. NMR data were complemented with molecular dynamics simulations and Corcema-ST to establish a 3D view on the molecular recognition process between Gal, GalNAc, and the Tn-presenting glycopeptides and MGL. Gal and GalNAc have a dual binding mode with opposite trend of the main interaction pattern and the differences in affinity can be explained by additional hydrogen bonds and CH-p contacts involving exclusively the NHAc moiety.

AB - The human macrophage galactose-type lectin (MGL) is a key physiological receptor for the carcinoma-associated Tn antigen (GalNAc-α-1-O-Ser/Thr) in mucins. NMR and modeling-based data on the molecular recognition features of synthetic Tn-bearing glycopeptides by MGL are presented. Cognate epitopes on the sugar and matching key amino acids involved in the interaction were identified by saturation transfer difference (STD) NMR spectroscopy. Only the amino acids close to the glycosylation site in the peptides are involved in lectin contact. Moreover, control experi-ments with non-glycosylated MUC1 peptides unequivocally showed that the sugar residue is essential for MGL binding, as is Ca2+. NMR data were complemented with molecular dynamics simulations and Corcema-ST to establish a 3D view on the molecular recognition process between Gal, GalNAc, and the Tn-presenting glycopeptides and MGL. Gal and GalNAc have a dual binding mode with opposite trend of the main interaction pattern and the differences in affinity can be explained by additional hydrogen bonds and CH-p contacts involving exclusively the NHAc moiety.

KW - Binding studies

KW - Macrophage galactose-type lectin

KW - Molecular modeling

KW - Molecular recognition

KW - Mucin glycopeptides

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DO - 10.1002/chem.201404566

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