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Intra- and inter-molecular interactions of human galectin-3: assessment by full-assignment-based NMR.

Ippel, Hans; Miller, Michelle C; Vértesy, Sabine; Zhang, Yi; Cañada, F Javier; Suylen, Dennis; Umemoto, Kimiko; Romanò, Cecilia; Hackeng, Tilman and Tai, Guihua, et al. (2016) In Glycobiology 26(8). p.888-903
Abstract
Galectin-3 is an adhesion/growth-regulatory protein with a modular design comprising an N-terminal tail (NT, residues 1-111) and the conserved carbohydrate recognition domain (CRD, residues 112-250). The chimera-type galectin interacts with both glycan and peptide motifs. Complete (13)C/(15)N-assignment of the human protein makes NMR-based analysis of its structure beyond the CRD possible. Using two synthetic NT polypeptides covering residues 1-50 and 51-107, evidence for transient secondary structure was found with helical conformation from residues 5 to 15 as well as proline-mediated, multi-turn structure from residues 18 to 32 and around PGAYP repeats. Intramolecular interactions occur between the CRD F-face (the 5-stranded β-sheet... (More)
Galectin-3 is an adhesion/growth-regulatory protein with a modular design comprising an N-terminal tail (NT, residues 1-111) and the conserved carbohydrate recognition domain (CRD, residues 112-250). The chimera-type galectin interacts with both glycan and peptide motifs. Complete (13)C/(15)N-assignment of the human protein makes NMR-based analysis of its structure beyond the CRD possible. Using two synthetic NT polypeptides covering residues 1-50 and 51-107, evidence for transient secondary structure was found with helical conformation from residues 5 to 15 as well as proline-mediated, multi-turn structure from residues 18 to 32 and around PGAYP repeats. Intramolecular interactions occur between the CRD F-face (the 5-stranded β-sheet behind the canonical carbohydrate-binding 6-stranded β-sheet of the S-face) and NT in full-length galectin-3, with the sequence P(23)GAW(26) … P(37)GASYPGAY(45) defining the primary binding epitope within the NT. Work with designed peptides indicates that the PGAX motif is crucial for self-interactions between NT/CRD. Phosphorylation at position Ser6 (and Ser12) (a physiological modification) and the influence of ligand binding have minimal effect on this interaction. Lastly, galectin-3 molecules can interact weakly with each other via the F-faces of their CRDs, an interaction that appears to be assisted by their NTs. Overall, our results add insight to defining binding sites on galectin-3 beyond the canonical contact area for β-galactosides. (Less)
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Glycobiology
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26
issue
8
pages
16 pages
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Oxford University Press
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  • pmid:26911284
  • scopus:84988535559
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1460-2423
DOI
10.1093/glycob/cww021
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English
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cb8fd38f-9f6c-4249-adf4-08020a03552f (old id 8821758)
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2016-03-03 13:18:07
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@article{cb8fd38f-9f6c-4249-adf4-08020a03552f,
  abstract     = {Galectin-3 is an adhesion/growth-regulatory protein with a modular design comprising an N-terminal tail (NT, residues 1-111) and the conserved carbohydrate recognition domain (CRD, residues 112-250). The chimera-type galectin interacts with both glycan and peptide motifs. Complete (13)C/(15)N-assignment of the human protein makes NMR-based analysis of its structure beyond the CRD possible. Using two synthetic NT polypeptides covering residues 1-50 and 51-107, evidence for transient secondary structure was found with helical conformation from residues 5 to 15 as well as proline-mediated, multi-turn structure from residues 18 to 32 and around PGAYP repeats. Intramolecular interactions occur between the CRD F-face (the 5-stranded β-sheet behind the canonical carbohydrate-binding 6-stranded β-sheet of the S-face) and NT in full-length galectin-3, with the sequence P(23)GAW(26) … P(37)GASYPGAY(45) defining the primary binding epitope within the NT. Work with designed peptides indicates that the PGAX motif is crucial for self-interactions between NT/CRD. Phosphorylation at position Ser6 (and Ser12) (a physiological modification) and the influence of ligand binding have minimal effect on this interaction. Lastly, galectin-3 molecules can interact weakly with each other via the F-faces of their CRDs, an interaction that appears to be assisted by their NTs. Overall, our results add insight to defining binding sites on galectin-3 beyond the canonical contact area for β-galactosides.},
  author       = {Ippel, Hans and Miller, Michelle C and Vértesy, Sabine and Zhang, Yi and Cañada, F Javier and Suylen, Dennis and Umemoto, Kimiko and Romanò, Cecilia and Hackeng, Tilman and Tai, Guihua and Leffler, Hakon and Kopitz, Jürgen and André, Sabine and Kübler, Dieter and Jiménez-Barbero, Jesús and Oscarson, Stefan and Gabius, Hans-Joachim and Mayo, Kevin H},
  issn         = {1460-2423},
  language     = {eng},
  month        = {02},
  number       = {8},
  pages        = {888--903},
  publisher    = {Oxford University Press},
  series       = {Glycobiology},
  title        = {Intra- and inter-molecular interactions of human galectin-3: assessment by full-assignment-based NMR.},
  url          = {http://dx.doi.org/10.1093/glycob/cww021},
  volume       = {26},
  year         = {2016},
}