• J. Biol. Chem. · Apr 2015

    HypE-specific nanobodies as tools to modulate HypE-mediated target AMPylation.

    • Matthias C Truttmann, Qin Wu, Sarah Stiegeler, Joao N Duarte, Jessica Ingram, and Hidde L Ploegh.
    • From the Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142 and.
    • J. Biol. Chem. 2015 Apr 3; 290 (14): 9087-100.

    AbstractThe covalent addition of mono-AMP to target proteins (AMPylation) by Fic domain-containing proteins is a poorly understood, yet highly conserved post-translational modification. Here, we describe the generation, evaluation, and application of four HypE-specific nanobodies: three that inhibit HypE-mediated target AMPylation in vitro and one that acts as an activator. All heavy chain-only antibody variable domains bind HypE when expressed as GFP fusions in intact cells. We observed localization of HypE at the nuclear envelope and further identified histones H2-H4, but not H1, as novel in vitro targets of the human Fic protein. Its role in histone modification provides a possible link between AMPylation and regulation of gene expression.© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

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