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Glycoconjugate Journal

, Volume 34, Issue 3, pp 405–410 | Cite as

19F labelled glycosaminoglycan probes for solution NMR and non-linear (CARS) microscopy

  • Marcelo A. Lima
  • Renan P. Cavalheiro
  • Gustavo M.Viana
  • Maria C.Z. Meneghetti
  • Timothy R. Rudd
  • Mark A. Skidmore
  • Andrew K. Powell
  • Edwin A. YatesEmail author
Original Article

Abstract

Studying polysaccharide-protein interactions under physiological conditions by conventional techniques is challenging. Ideally, macromolecules could be followed by both in vitro spectroscopy experiments as well as in tissues using microscopy, to enable a proper comparison of results over these different scales but, often, this is not feasible. The cell surface and extracellular matrix polysaccharides, glycosaminoglycans (GAGs) lack groups that can be detected selectively in the biological milieu. The introduction of 19F labels into GAG polysaccharides is explored and the interaction of a labelled GAG with the heparin-binding protein, antithrombin, employing 19F NMR spectroscopy is followed. Furthermore, the ability of 19F labelled GAGs to be imaged using CARS microscopy is demonstrated. 19F labelled GAGs enable both 19F NMR protein-GAG binding studies in solution at the molecular level and non-linear microscopy at a microscopic scale to be conducted on the same material, essentially free of background signals.

Keywords

Heparin NMR 19Non-linear microscopy 

Notes

Acknowledgments

MAL and EAY gratefully acknowledge Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for financial support. The authors also gratefully acknowledge Dr. Tony Curtis of Keele University for the provision of 19F NMR spectra. Dr. Andrew V. Stachulski of the Department of Chemistry, University of Liverpool is thanked for useful discussions and advice. The authors also thank Dr. Claudio Tormena of University of Campinas (UNICAMP), Brazil for provision of NMR facilities.

Compliance with ethical standards

Conflicts of interest

The authors declare that they have no conflicts of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

10719_2016_9723_MOESM1_ESM.docx (179 kb)
Table S1 (DOCX 178 kb)
10719_2016_9723_MOESM2_ESM.docx (112 kb)
Table S2 (DOCX 112 kb)

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Marcelo A. Lima
    • 1
    • 2
  • Renan P. Cavalheiro
    • 1
  • Gustavo M.Viana
    • 1
  • Maria C.Z. Meneghetti
    • 1
  • Timothy R. Rudd
    • 3
    • 2
  • Mark A. Skidmore
    • 2
    • 4
  • Andrew K. Powell
    • 5
    • 2
  • Edwin A. Yates
    • 2
    • 1
    • 4
    Email author
  1. 1.Department of BiochemistryUNIFESPSão PauloBrazil
  2. 2.Department of BiochemistryUniversity of LiverpoolLiverpoolUK
  3. 3.The National Institute of Biological Standards and Controls, Blanche LanePotters BarUK
  4. 4.School of Life SciencesKeele UniversityKeeleUK
  5. 5.School of Pharmacy and Biomolecular SciencesLiverpool John Moores UniversityLiverpoolUK

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