Biotinylated anisomycin: A comparison of classical and "click" chemistry approaches

Iain A. Inverarity, Romain F H Viguier, Philip Cohen, Alison N. Hulme

    Research output: Contribution to journalArticle

    17 Citations (Scopus)

    Abstract

    Two approaches to the synthesis of biotinylated derivatives of the stress-activated protein kinase (SAPK) pathway activator anisomycin have been investigated. Attachment of the biotin moiety to the central core was achieved either through the use of a classical displacement reaction on α-halo carbonyl derivatives of biotin or through a copper(I)-catalyzed 1,3-dipolar Huisgen cycloaddition ("click") coupling of biotinylated azides to propargyl-marked analogues of anisomycin. In each case, the resultant N-linked molecular probes were found to be active in SAPK pathway immunoblot assays, while their O-linked counterparts were inactive. However, in sharp contrast to the classical coupling approach which results in low coupling yields, the aqueous "click" coupling process was found to deliver high yields of biotinylated probes, making it the conjugation method of choice. A survey of the available methods for the addition of a propargyl marker onto a range of chemical functionalities strongly suggests that this copper(I)-catalyzed 1,3-dipolar Huisgen cycloaddition approach to biotinylation may be generally applied.

    Original languageEnglish
    Pages (from-to)1593-1603
    Number of pages11
    JournalBioconjugate Chemistry
    Volume18
    Issue number5
    DOIs
    Publication statusPublished - 1 Sep 2007

    Fingerprint

    Anisomycin
    Click Chemistry
    Cycloaddition
    Cycloaddition Reaction
    Biotin
    Heat-Shock Proteins
    Protein Kinases
    Copper
    Biotinylation
    Derivatives
    Proteins
    Molecular Probes
    Azides
    Assays
    Surveys and Questionnaires

    Cite this

    Inverarity, Iain A. ; Viguier, Romain F H ; Cohen, Philip ; Hulme, Alison N. / Biotinylated anisomycin : A comparison of classical and "click" chemistry approaches. In: Bioconjugate Chemistry. 2007 ; Vol. 18, No. 5. pp. 1593-1603.
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    abstract = "Two approaches to the synthesis of biotinylated derivatives of the stress-activated protein kinase (SAPK) pathway activator anisomycin have been investigated. Attachment of the biotin moiety to the central core was achieved either through the use of a classical displacement reaction on α-halo carbonyl derivatives of biotin or through a copper(I)-catalyzed 1,3-dipolar Huisgen cycloaddition ({"}click{"}) coupling of biotinylated azides to propargyl-marked analogues of anisomycin. In each case, the resultant N-linked molecular probes were found to be active in SAPK pathway immunoblot assays, while their O-linked counterparts were inactive. However, in sharp contrast to the classical coupling approach which results in low coupling yields, the aqueous {"}click{"} coupling process was found to deliver high yields of biotinylated probes, making it the conjugation method of choice. A survey of the available methods for the addition of a propargyl marker onto a range of chemical functionalities strongly suggests that this copper(I)-catalyzed 1,3-dipolar Huisgen cycloaddition approach to biotinylation may be generally applied.",
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    Biotinylated anisomycin : A comparison of classical and "click" chemistry approaches. / Inverarity, Iain A.; Viguier, Romain F H; Cohen, Philip; Hulme, Alison N.

    In: Bioconjugate Chemistry, Vol. 18, No. 5, 01.09.2007, p. 1593-1603.

    Research output: Contribution to journalArticle

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    AU - Viguier, Romain F H

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