Peptide biotinylation with amine-reactive esters: Differential side chain reactivity

Brian T. Miller, Thomas J. Collins, Mark E. Rogers, Alexander Kurosky

    Research output: Contribution to journalArticle

    36 Citations (Scopus)

    Abstract

    N-hydroxysuccinimide (NHS) esters of biotin are reported to react specifically with amino groups of peptides and proteins. However, we have found that these reagents can readily acylate other functional groups in specific peptide sequences under relatively mild conditions. We have extended our inquiry of sequence-dependent acylation by evaluating the reactivity of a variety of commonly employed biotinylation reagents typically used for amino group modification. These included the p-nitrophenyl ester of biotin, NHS- esters of biotin containing aminohexanoic acid spacer arms, and a sulfonated NHS-biotin ester that contained a disulfide bond within its spacer. The decapeptide [D-Lys6]gonadotropin releasing hormone was employed as a model peptide. Reaction products were characterized by high-performance liquid chromatography, amino acid compositional analysis, reaction with hydroxylamine, and mass spectrometry. In addition to the O-acylation of Ser4 and Tyr5 in this peptide, we have also identified a novel biotinylation of the Arg8 side chain.

    Original languageEnglish (US)
    Pages (from-to)1585-1595
    Number of pages11
    JournalPeptides
    Volume18
    Issue number10
    DOIs
    StatePublished - 1997

    Fingerprint

    Biotinylation
    Amines
    Esters
    Biotin
    Acylation
    Peptides
    Aminocaproates
    Hydroxylamine
    High performance liquid chromatography
    Reaction products
    Gonadotropin-Releasing Hormone
    Disulfides
    Functional groups
    Mass spectrometry
    Mass Spectrometry
    High Pressure Liquid Chromatography
    Amino Acids
    N-hydroxysuccinimide
    Proteins

    Keywords

    • Aminohexanoic acid
    • Arginine modification
    • Biotin-p-nitrophenyl ester
    • Biotinylation [D-Lys]GnRH
    • NHS-biotin esters
    • O-acylation
    • Solvent effects

    ASJC Scopus subject areas

    • Biochemistry
    • Endocrinology
    • Physiology
    • Cellular and Molecular Neuroscience

    Cite this

    Peptide biotinylation with amine-reactive esters : Differential side chain reactivity. / Miller, Brian T.; Collins, Thomas J.; Rogers, Mark E.; Kurosky, Alexander.

    In: Peptides, Vol. 18, No. 10, 1997, p. 1585-1595.

    Research output: Contribution to journalArticle

    Miller, BT, Collins, TJ, Rogers, ME & Kurosky, A 1997, 'Peptide biotinylation with amine-reactive esters: Differential side chain reactivity', Peptides, vol. 18, no. 10, pp. 1585-1595. https://doi.org/10.1016/S0196-9781(97)00225-8
    Miller, Brian T. ; Collins, Thomas J. ; Rogers, Mark E. ; Kurosky, Alexander. / Peptide biotinylation with amine-reactive esters : Differential side chain reactivity. In: Peptides. 1997 ; Vol. 18, No. 10. pp. 1585-1595.
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    AU - Kurosky, Alexander

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    KW - Solvent effects

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