Attachment of peptide building blocks to proteins through tyrosine bioconjugation.
Attachment of peptide building blocks to proteins through tyrosine bioconjugation.
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DOI：
10.1021/bc700231v
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发表时间：
2008
影响因子：
4.7
通讯作者：
Dante W. Romanini;M. Francis
中科院分区：
文献类型：
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作者：
Dante W. Romanini;M. Francis
最近的努力已经产生了许多具有有用的结合、传感和细胞摄取特性的短肽序列。为了将这些序列连接到完整蛋白质上的酪氨酸残基上，报道了三组分曼尼希型策略。开发了两种固相合成路线来获得长度长达 20 个残基且 N 或 C 末端带有苯胺的肽。在 20 mM 甲醛存在下，这些官能团在温和的反应条件下与蛋白质上的酪氨酸残基偶联。使用质谱和免疫印迹分析证实了所得生物缀合物的身份。筛选实验证明该方法与含有所有氨基酸（包括赖氨酸和半胱氨酸残基）的底物兼容。重要的是，蛋白质上的酪氨酸残基表现出更快的反应速率，使得含有该残基的短肽可以在没有交叉反应的情况下进行偶联。
Recent efforts have yielded a number of short peptide sequences with useful binding, sensing, and cellular uptake properties. In order to attach these sequences to tyrosine residues on intact proteins, a three-component Mannich-type strategy is reported. Two solid phase synthetic routes were developed to access peptides up to 20 residues in length with anilines at either the N- or C-termini. In the presence of 20 mM formaldehyde, these functional groups were coupled to tyrosine residues on proteins under mild reaction conditions. The identities of the resulting bioconjugates were confirmed using mass spectrometry and immunoblot analysis. Screening experiments have demonstrated that the method is compatible with substrates containing all of the amino acids, including lysine and cysteine residues. Importantly, tyrosine residues on proteins exhibit much faster reaction rates, allowing short peptides containing this residue to be coupled without cross reactions.