title: 19F labelled glycosaminoglycan probes for solution NMR and non-linear (CARS) microscopy.
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 of19F labels into GAG polysaccharides is explored and the interaction of a labelled GAG with the heparin-binding protein, antithrombin, employing19F NMR spectroscopy is followed. Furthermore, the ability of19F labelled GAGs to be imaged using CARS microscopy is demonstrated.19F labelled GAGs enable both19F 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.
authors: Department of Biochemistry, UNIFESP, Rua Três de Maio, Vila Clementino, São Paulo, SP, 40440, Brazil.|Department of Biochemistry, University of Liverpool, L69 7ZB, Liverpool, UK.
authors: Marcelo A
authors: MA
authors: Lima
authors: Department of Biochemistry, UNIFESP, Rua Três de Maio, Vila Clementino, São Paulo, SP, 40440, Brazil.
authors: Renan P
authors: RP
authors: Cavalheiro
authors: Department of Biochemistry, UNIFESP, Rua Três de Maio, Vila Clementino, São Paulo, SP, 40440, Brazil.
authors: Gustavo
authors: G
authors: M Viana
authors: Department of Biochemistry, UNIFESP, Rua Três de Maio, Vila Clementino, São Paulo, SP, 40440, Brazil.
authors: Maria C Z
authors: MCZ
authors: Meneghetti
authors: Department of Biochemistry, University of Liverpool, L69 7ZB, Liverpool, UK.|The National Institute of Biological Standards and Controls, Blanche Lane, South Mimms, Potters Bar, Hertfordshire, EN6 3QG, UK.
authors: Timothy R
authors: TR
authors: Rudd
authors: Department of Biochemistry, University of Liverpool, L69 7ZB, Liverpool, UK.|School of Life Sciences, Keele University, Keele, Staffordshire, ST5 5BG, UK.
authors: Mark A
authors: MA
authors: Skidmore
authors: Department of Biochemistry, University of Liverpool, L69 7ZB, Liverpool, UK.|School of Pharmacy and Biomolecular Sciences, Liverpool John Moores University, L3 3AF, Liverpool, UK.
authors: Andrew K
authors: AK
authors: Powell
authors: Department of Biochemistry, UNIFESP, Rua Três de Maio, Vila Clementino, São Paulo, SP, 40440, Brazil. eayates@liv.ac.uk.|Department of Biochemistry, University of Liverpool, L69 7ZB, Liverpool, UK. eayates@liv.ac.uk.|School of Life Sciences, Keele University, Keele, Staffordshire, ST5 5BG, UK. eayates@liv.ac.uk.
authors: Edwin A
authors: EA
authors: Yates
mesh_terms: D000107:Acetylation; D000991:Antithrombins / Q000737:chemistry; D005461:Fluorine / Q000737:chemistry*; D066235:Fluorine-19 Magnetic Resonance Imaging / Q000379:methods*; D006025:Glycosaminoglycans / Q000032:analysis / Q000737:chemistry*; D054879:Halogenation; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D015335:Molecular Probes / Q000032:analysis / Q000737:chemistry*; D011485:Protein Binding; D012996:Solutions; D013059:Spectrum Analysis, Raman / Q000379:methods; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D000991:Antithrombins; D006025:Glycosaminoglycans; D015335:Molecular Probes; D012996:Solutions; D005461:Fluorine
keywords: 19F; Heparin; NMR; Non-linear microscopy