Tyrosinase-Mediated Bioconjugation. A Versatile Approach to Chimeric Macromolecules
Tyrosinase-Mediated Bioconjugation. A Versatile Approach to Chimeric Macromolecules
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DOI：
10.1021/acs.bioconjchem.8b00227
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发表时间：
2018-08-01
影响因子：
4.7
通讯作者：
Tirelli, Nicola
中科院分区：
文献类型：
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作者：
Montanari, Elita;Gennari, Arianna;Tirelli, Nicola
提出了一种酪氨酸选择性可逆生物偶联方法；酪氨酸被酶转化为儿茶酚，并在原位“点击”到硼酸上。重要的是，我们的工艺选择性地产生儿茶酚，避免醌，从而改善了对产品化学特性的控制。我们将含玻尿酸（HyA）的硼酸偶联到含有不同数量和位置的酪氨酸的肽上；使用标记肽来提供暴露良好的酪氨酸残基，在我们的情况下，血凝素衍生的ha标签使我们的方法适用于几乎任何蛋白质；我们通过将ha标记的卵清蛋白偶联到HyA上证明了这一概念，从而也证明了生产嵌合蛋白聚糖的可行性。这种方法需要注意的是，尽管硼酯的形成不会影响底物（卵清蛋白和HyA）的生物学识别，但儿茶酚的引入可能会改变它们的一些生物学特性：例如，只有在酪氨酸酶处理后，卵清蛋白才能直接诱导树突状细胞成熟，无论是单独的还是作为HyA缀合物。
We present a method for tyrosine-selective and reversible bioconjugation; tyrosines are enzymatically converted into catechols and in situ "clicked" onto boronic acids. Importantly, our process selectively produces catechols and avoids quinones, thereby improving the control over the chemical identity of the products. We have conjugated boronic acid containing hyaluronic acid (HyA) to peptides bearing tyrosines in variable number and position; the use of tagging peptides for the provision of well exposed tyrosine residues in our case the hemagglutinin-derived HA-tag makes our approach applicable to virtually any protein; we have demonstrated this concept by conjugating HA-tagged ovalbumin to HyA, thereby also showing the feasibility of producing chimeric proteoglycans. A caveat of this appproach is that, although the formation of boronic esters does not affect the biological recognition of substrates (ovalbumin and HyA), the introduction of catechols may alter some of their biological properties: for example, only after tyrosinase treatment ovalbumin directly induced dendritic cell maturation, either alone or as a HyA conjugate.