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A Stable Pyrophosphoserine Analog for Incorporation into Peptides and Proteins
journal contribution
posted on 2016-02-05, 20:49 authored by Lisa M. Yates, Dorothea FiedlerProtein
pyrophosphorylation is a covalent modification of proteins,
mediated by the inositol pyrophosphate messengers. Although the inositol
pyrophosphates have been linked to a range of cellular processes,
the role of protein pyrophosphorylation remains minimally characterized in vivo. The inherent instability of the phosphoanhydride
bond has hampered the development of useful bioanalytical techniques
to interrogate this novel signaling mechanism. Here, we describe the
preparation of a pyrophosphoserine analog containing a stable methylene-bisphosphonate
group that is compatible with solid-phase peptide synthesis. The resulting
peptides demonstrate enhanced stability in Eukaryotic cell lysates
and mammalian plasma and display resistance toward chemical degradation,
when compared to the corresponding pyrophosphopeptides. In addition,
the peptides containing the stable pyrophosphoserine analog are highly
compatible with common ligation methods, such as native chemical ligation,
maleimide conjugation, and glutaraldehyde ligation. The bisphosphonate-containing
peptides will, therefore, be well-suited for future pyrophosphoserine
antibody generation and affinity capture of pyrophosphoprotein binding
partners and provide a key entry point to study the regulatory role
of protein pyrophosphorylation.