- A Stable Pyrophosphoserine Analog for Incorporation into Peptides and Proteins
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Protein 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.
- Yates, Lisa M.,Fiedler, Dorothea
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- Preparations of Nα-Fmoc-O-[(benzyloxy)hydroxyphosphinyl] β-hydroxy α-amino acid derivatives
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Nα-Fmoc-O-[(benzyloxy)hydroxyphosphinyl]serine and Nα-Fmoc-O-[(benzyloxy)hydroxyphosphinyl]threonine were prepared in order to be utilized for the synthesis of phosphoserine- or phosphothreonine-containing peptides based on the Fmoc-mode pre-phosphorylation strategy. These derivatives were obtained as crystalline compounds, which are favorable for use as building blocks for an automated peptide synthesis by a solid-phase method.
- Wakamiya, Tateaki,Nishida, Takatoshi,Togashi, Ryusaku,Saruta, Kunio,Yasuoka, Jun-Ichi,Kusumoto, Shoichi
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p. 465 - 468
(2007/10/03)
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- An Efficient Procedure for Solid Phase Synthesis of Phosphopeptides by the Fmoc Strategy
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The Fmoc-mode solid-phase method was succesfully applied to establish a practical procedure for the synthesis of phosphopeptides.Both Fmoc-phosphoserine with free phosphoric moiety and its monobenzyl phosphate derivative were examined as one of starting materials for the synthesis of peptides.The employment of the latter gave better result in respects of the yield and purity of the product than that obtained with the former.
- Wakamiya, Tateaki,Saruta, Kunio,Yasuoka, Jun-ichi,Kusumoto, Shoichi
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p. 1099 - 1102
(2007/10/02)
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- Application of t-Butyldimethylsilyl Ethers of Serine, Threonine and Tyrosine in Peptide Synthesis
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The utility of Tbdms (t-butyldimethylsilyl) ethers, prepared conveniently in a one pot procedure from Nα-Fmoc (9-fluorenylmethoxycarbonyl) and Nα-Z (benzyloxycarbonyl) hydroxyamino acids, is demonstrated: peptide bond formation and esterification to 4-alkoxybenzylalcohol resin are achieved readily with these derivatives.The lability of the Tbdms ethers to various reagents enables selective deprotection of the hydroxyl side-chains assembly, desirable, e.g., for phosphorylation of glycosylation.
- Fischer, Peter M.
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p. 7605 - 7608
(2007/10/02)
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