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Fluoroglycoproteins by Copper-Free Strain-Promoted Azide–Alkyne Cycloaddition

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Peptide and Protein Engineering

Part of the book series: Springer Protocols Handbooks ((SPH))

Abstract

This chapter describes a reliable two-step, metal-free protocol for the preparation of well-defined fluoroglycoproteins. It starts with a first alkylation step to chemoselectively install strained alkyne handles at cysteine residues followed by a second strain-promoted azide–alkyne cycloaddition using an inverse electron-demand Diels–Alder reaction. This proof-of-principle study that uses the apoptotic protein marker Annexin V enables the efficient metal-free incorporation of 2-deoxy-2-fluoro-glycopyranosyl azides into proteins and complements previous methods using Cu(I)-mediated azide–alkyne cycloadditions and thiol chemistry.

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Abbreviations

ADIBO:

Azadibenzobicyclooctyne

AnxV:

Annexin V

BCN:

Bicyclo[6.1.0]nonyne

CuAAC:

Copper-mediated azide–alkyne cycloaddition

DBCO:

Dibenzocyclooctyne

DIFO:

Difluorinated cyclooctyne

DMF:

N,N-Dimethylformamide

ESI:

Electrospray ionization

FDG:

2-Deoxy-2-fluoro-β-d-glucopyranose

FDGN3:

2-Deoxy-2-fluoro-β-d-glucopyranosyl azide

HPLC:

High-performance liquid chromatography

LC:

Liquid chromatography

MS:

Mass spectrometry

MWCO:

Molecular weight cut-off

SPAAC:

Strain-promoted azide–alkyne cycloaddition

TOF:

Time of flight

UPLC:

Ultra-performance liquid chromatography

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Acknowledgments

The authors thank the Spanish Government (MCIU) (CTQ2017-90088-R to O.B.), the national agency of investigation (AEI), the European Regional Development Fund, the Royal Society (URF to G.J.L.B., URF/R/180019), FCT Portugal (iFCT to G.J.L.B., IF/00624/2015 and postdoctoral fellowship to P.M.S.D.C., SFRH/BPD/103172/2014), and the EPSRC (EP/M003647/1 to G.J.L.B.) for financial support. O.B. is a Ramón y Cajal Fellow (RYC-2015-17705).

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Authors and Affiliations

  1. Department of Chemistry, University of Cambridge, Cambridge, UK

    Pedro M. S. D. Cal & Gonçalo J. L. Bernardes

  2. Instituto de Medicina Molecular, Universidade de Lisboa, Lisboa, Portugal

    Pedro M. S. D. Cal & Gonçalo J. L. Bernardes

  3. Departament de Química Analítica i Química Orgànica, Universitat Rovira i Virgili, Tarragona, Spain

    Omar Boutureira

Authors
  1. Pedro M. S. D. Cal
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  2. Gonçalo J. L. Bernardes
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  3. Omar Boutureira
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Corresponding author

Correspondence to Omar Boutureira .

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Editors and Affiliations

  1. iSm2 UMR CNRS 7313, Aix-Marseille Université, Marseille, France

    Olga Iranzo

  2. UCIBIO, Chemistry Department, School of Science and Technology, NOVA University of Lisbon, Caparica, Portugal

    Ana Cecília Roque

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Cal, P.M.S.D., Bernardes, G.J.L., Boutureira, O. (2020). Fluoroglycoproteins by Copper-Free Strain-Promoted Azide–Alkyne Cycloaddition. In: Iranzo, O., Roque, A. (eds) Peptide and Protein Engineering. Springer Protocols Handbooks. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0720-6_5

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  • DOI: https://doi.org/10.1007/978-1-0716-0720-6_5

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0719-0

  • Online ISBN: 978-1-0716-0720-6

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