The generality of kinase-catalyzed biotinylation.
The generality of kinase-catalyzed biotinylation.
复制标题
激酶催化生物素化的一般性。
DOI：
10.1016/j.bmc.2015.11.029
复制
发表时间：
2016
影响因子：
3.5
通讯作者：
Pflum,MaryKayH
中科院分区：
文献类型：
--
作者：
Senevirathne,Chamara;Embogama,DMaheeka;Anthony,ThilaniA;Fouda,AhmedE;Pflum,MaryKayH
Kinase-catalyzed protein phosphorylation is involved in a wide variety of cellular events. Development of methods to monitor phosphoproteins in normal and diseased states is critical to fully characterize cell signaling. Towards phosphoprotein analysis tools, our lab reported kinase-catalyzed labeling where γ-phosphate modified ATP analogs are utilized by kinases to label peptides or protein substrates with a functional tag. In particular, the ATP-biotin analog was developed for kinase-catalyzed biotinylation. However, kinase-catalyzed labeling has been tested rigorously with only a few kinases, preventing use of ATP-biotin as a general tool. Here, biotinylation experiments, gel or HPLC-based quantification, and kinetic measurements indicated that twenty-five kinases throughout the kinome tree accepted ATP-biotin as a cosubstrate. With this rigorous characterization of ATP-biotin compatibility, kinase-catalyzed labeling is now immediately useful for studying phosphoproteins and characterizing the role of phosphorylation in various biological events.
