Diversification of polyphosphate end-labeling via bridging molecules
Diversification of polyphosphate end-labeling via bridging molecules
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DOI：
10.1371/journal.pone.0237849
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发表时间：
2020-08-21
期刊：
影响因子：
3.7
通讯作者：
Morrissey, James H.
中科院分区：
文献类型：
--
作者：
Baker, Catherine J.;Smith, Stephanie A.;Morrissey, James H.
我们之前开发了一种基于碳二亚胺的方法，将含有伯胺的分子共价偶联到聚磷酸盐的末端磷酸盐上，从而促进了对无机聚磷酸盐生物学作用的研究。现在，我们通过优化反应条件并使用容易获得的包含伯胺和附加反应部分(包括另一个伯胺、硫醇或点击化学试剂，如二苯并环辛基)的“桥联分子”来扩展该方法。这种两步标记法用于将商业上可获得的生物素、肽表位标签和荧光染料的衍生物共价连接到聚磷酸盐的末端磷酸盐上。此外，我们还报道了三种从过量反应物中提纯共轭聚磷酸盐的简便方法。
Investigation of the biological roles of inorganic polyphosphate has been facilitated by our previous development of a carbodiimide-based method for covalently coupling primary amine-containing molecules to the terminal phosphates of polyphosphate. We now extend that approach by optimizing the reaction conditions and using readily available "bridging molecules" containing a primary amine and an additional reactive moiety, including another primary amine, a thiol or a click chemistry reagent such as dibenzocyclooctyne. This two-step labeling method is used to covalently attach commercially available derivatives of biotin, peptide epitope tags, and fluorescent dyes to the terminal phosphates of polyphosphate. Additionally, we report three facile methods for purifying conjugated polyphosphate from excess reactants.