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
UR - http://www.scopus.com/inward/record.url?scp=84915746309&partnerID=8YFLogxK
U2 - 10.1002/chem.201404566
DO - 10.1002/chem.201404566
M3 - Article
C2 - 25346263
AN - SCOPUS:84915746309
SN - 0947-6539
VL - 20
SP - 16147
EP - 16155
JO - Chemistry - A European Journal
JF - Chemistry - A European Journal
IS - 49
ER -