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

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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.

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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.

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Authors and Affiliations

  1. Department of Biochemistry, UNIFESP, Rua Três de Maio, Vila Clementino, São Paulo, SP, 40440, Brazil

    Marcelo A. Lima, Renan P. Cavalheiro, Gustavo M.Viana, Maria C.Z. Meneghetti & Edwin A. Yates

  2. Department of Biochemistry, University of Liverpool, L69 7ZB, Liverpool, UK

    Marcelo A. Lima, Timothy R. Rudd, Mark A. Skidmore, Andrew K. Powell & Edwin A. Yates

  3. The National Institute of Biological Standards and Controls, Blanche Lane, South Mimms, Potters Bar, Hertfordshire, EN6 3QG, UK

    Timothy R. Rudd

  4. School of Life Sciences, Keele University, Keele, Staffordshire, ST5 5BG, UK

    Mark A. Skidmore & Edwin A. Yates

  5. School of Pharmacy and Biomolecular Sciences, Liverpool John Moores University, L3 3AF, Liverpool, UK

    Andrew K. Powell

Authors
  1. Marcelo A. Lima
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  2. Renan P. Cavalheiro
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  3. Gustavo M.Viana
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  4. Maria C.Z. Meneghetti
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  5. Timothy R. Rudd
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  6. Mark A. Skidmore
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  7. Andrew K. Powell
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  8. Edwin A. Yates
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Correspondence to Edwin A. Yates.

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

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Lima, M.A., Cavalheiro, R.P., M.Viana, G. et al. 19F labelled glycosaminoglycan probes for solution NMR and non-linear (CARS) microscopy. Glycoconj J 34, 405–410 (2017). https://doi.org/10.1007/s10719-016-9723-x

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  • DOI: https://doi.org/10.1007/s10719-016-9723-x

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