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

Hans Ippel, Michelle C Miller, Sabine Vértesy, Yi Zhang, F Javier Cañada, Dennis Suylen, Kimiko Umemoto, Cecilia Romanò, Tilman Hackeng, Guihua Tai

Research output: Contribution to journalArticle

26 Citations (Scopus)

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.
Original languageEnglish (US)
Pages (from-to)1-16
Number of pages16
JournalGlycobiology
DOIs
StatePublished - 2016

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Galectin 3
Molecular interactions
Nuclear magnetic resonance
Peptides
Pyridinolcarbamate
Carbohydrates
Galectins
Galactosides
Phosphorylation
Proline
Polysaccharides
Conformations
Tail
Epitopes
Proteins
Adhesion
Binding Sites
Ligands
Molecules
human galectin-3

Keywords

  • adhesion
  • all rights reserved
  • apoptosis
  • com
  • for permissions
  • journals
  • lectin
  • oup
  • permissions
  • phosphorylation
  • please e-mail
  • press
  • published by oxford university
  • self-association
  • the author 2016

PubMed: MeSH publication types

  • Journal Article

Cite this

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; Tai, Guihua.

In: Glycobiology, 2016, p. 1-16.

Research output: Contribution to journalArticle

Ippel, H, Miller, MC, Vértesy, S, Zhang, Y, Cañada, FJ, Suylen, D, Umemoto, K, Romanò, C, Hackeng, T & Tai, G 2016, 'Intra- and inter-molecular interactions of human galectin-3 : assessment by full-assignment-based NMR', Glycobiology, pp. 1-16. https://doi.org/10.1093/glycob/cww021
Ippel, Hans ; Miller, Michelle C ; Vértesy, Sabine ; Zhang, Yi ; Cañada, F Javier ; Suylen, Dennis ; Umemoto, Kimiko ; Romanò, Cecilia ; Hackeng, Tilman ; Tai, Guihua. / Intra- and inter-molecular interactions of human galectin-3 : assessment by full-assignment-based NMR. In: Glycobiology. 2016 ; pp. 1-16.
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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.",
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AU - Cañada, F Javier

AU - Suylen, Dennis

AU - Umemoto, Kimiko

AU - Romanò, Cecilia

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AB - 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.

KW - adhesion

KW - all rights reserved

KW - apoptosis

KW - com

KW - for permissions

KW - journals

KW - lectin

KW - oup

KW - permissions

KW - phosphorylation

KW - please e-mail

KW - press

KW - published by oxford university

KW - self-association

KW - the author 2016

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