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    Aminoquinoline Fluorescent Labels Obstruct Efficient Removal ofN‑Glycan Core α(1−6) Fucose by Bovine Kidney α‑L‑Fucosidase (BKF)


    O'Flaherty, Roisin and Harbison, Aoife M. and Hanley, Philip J. and Taron, Christopher H. and Fadda, Elisa and Rudd, Pauline M. (2017) Aminoquinoline Fluorescent Labels Obstruct Efficient Removal ofN‑Glycan Core α(1−6) Fucose by Bovine Kidney α‑L‑Fucosidase (BKF). Journal of Proteome Research, 16. pp. 4237-4243. ISSN 1535-3893

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    Abstract

    Here we report evidence that new aminoquinoline N-glycan fluorescent labels interfere with the release of core α(1−6) fucose from N-glycans by bovine kidney α-L-fucosidase (BKF). BKF is a commonly employed exoglycosidase for core α(1−6) fucose determination. Molecular simulations of the bound and unbound Fuc-α(1−6)-GlcNAc, where GlcNAc is situated at the reducing end for all N-glycans, suggest that the reduced BKF activity may be due to a nonoptimal fit of the highest populated conformers in the BKF active binding site at room temperature.Population analysis and free energy estimates suggest that an enhanced flexibility of the labeled sugar, which facilitates recognition and binding, can be achievable with extended reaction conditions. We provide these experimental conditions using a sequential exoglycosidase digestion array using high concentrations of BKF.

    Item Type: Article
    Keywords: aminoquinoline (AQC;) bovine kidney fucosidase (BKF); N-glycan ultra performance; liquid chromatography (UPLC);
    Academic Unit: Faculty of Science and Engineering > Chemistry
    Item ID: 11518
    Identification Number: https://doi.org/10.1021/acs.jproteome.7b00580
    Depositing User: Elisa Fadda
    Date Deposited: 30 Oct 2019 13:58
    Journal or Publication Title: Journal of Proteome Research
    Publisher: American Chemical Society
    Refereed: Yes
    URI:

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