Enzymatic Modification of N-Terminal Proline Residues Using Phenol Derivatives
Enzymatic Modification of N-Terminal Proline Residues Using Phenol Derivatives
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DOI：
10.1021/jacs.8b10845
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发表时间：
2019-03-06
影响因子：
15
通讯作者：
Francis, Matthew B.
中科院分区：
文献类型：
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作者：
Maza, Johnathan C.;Bader, Daniel L. V.;Francis, Matthew B.
报道了一种方便的酶促策略，用于修饰蛋白质N-末端位置的脯氨酸残基。使用从双孢蘑菇（abTYR）中分离的酪氨酸酶，酚和儿茶酚被氧化成高活性的邻醌中间体，然后以高产率偶联到N-末端脯氨酸残基。这种生物缀合方法的主要优点包括（1）使用空气稳定的前体，如果需要，可以大规模制备，（2）温和的反应条件，包括低温，（3）靶向可以容易地引入大多数蛋白质上的天然官能团，和（4）使用分子氧作为唯一的氧化剂。这种偶联策略被成功地证明用于将各种苯酚衍生的货物分子附着到一系列蛋白质底物，包括自组装的病毒衣壳、酶和几丁质结合结构域（CBD）。CBD结合到酵母细胞表面的能力被发现不受这种修饰反应的干扰。
A convenient enzymatic strategy is reported for the modification of proline residues in the N-terminal positions of proteins. Using a tyrosinase enzyme isolated from Agaricus bisporus (abTYR), phenols and catechols are oxidized to highly reactive o-quinone intermediates that then couple to N-terminal proline residues in high yield. Key advantages of this bioconjugation method include (1) the use of air-stable precursors that can be prepared on large scale if needed, (2) mild reaction conditions, including low temperatures, (3) the targeting of native functional groups that can be introduced readily on most proteins, and (4) the use of molecular oxygen as the sole oxidant. This coupling strategy was successfully demonstrated for the attachment of a variety of phenol-derivatized cargo molecules to a series of protein substrates, including self-assembled viral capsids, enzymes, and a chitin binding domain (CBD). The ability of the CBD to bind to the surfaces of yeast cells was found to be unperturbed by this modification reaction.