Leiden Institute of Chemistry, Leiden University, PO Box 9502, 2300 RA Leiden, The Netherlands.
J Am Chem Soc. 2010 Apr 14;132(14):5236-40. doi: 10.1021/ja910940q.
Adenosine diphosphate ribosylation (ADP-ribosylation) is a widely occurring post-translational modification of proteins at nucleophilic side chains of amino acid residues, such as asparagine, glutamic acid, and arginine. Elucidation of the biological role of ADP-ribosylation events would benefit from the availability of well-defined ADP-ribosylated peptides. Main issues in the construction of synthetic ADP-ribosylated peptides involve the availability of protected ribosylated amino acids suitable for peptide synthesis, development of a protective group strategy for peptide fragments compatible with the integrity of the adenosine diphosphate moiety, and an efficient procedure for pyrophosphate formation. In this paper we present a first approach to the chemical synthesis of ADP-ribosylated peptides in solution and on solid support. We describe an efficient synthesis of suitably protected ribosylated asparagine and glutamine building blocks suitable for Fmoc-based peptide synthesis. We further demonstrate a successful application of these ribosylated amino acids in the assembly of three fully synthetic ADP-ribosylated peptides by solution and solid phase approaches.
腺苷二磷酸核糖基化 (ADP-ribosylation) 是一种广泛存在的蛋白质翻译后修饰，发生在天冬酰胺、谷氨酸和精氨酸等氨基酸残基的亲核侧链上。阐明 ADP-ribosylation 事件的生物学作用将受益于具有明确定义的 ADP-ribosylated 肽的可用性。合成 ADP-ribosylated 肽的构建中存在的主要问题涉及适用于肽合成的受保护的核糖基化氨基酸的可用性、开发与腺苷二磷酸部分完整性兼容的肽片段的保护基策略，以及焦磷酸形成的有效方法。本文我们提出了一种在溶液和固相中化学合成 ADP-ribosylated 肽的初步方法。我们描述了一种适用于 Fmoc 基肽合成的有效合成方法，得到了合适保护的核糖基化天冬酰胺和谷氨酰胺砌块。我们进一步证明了这些核糖基化氨基酸在通过溶液和固相方法组装三个完全合成的 ADP-ribosylated 肽中的成功应用。
