Global analysis of protein tyrosine phosphatase activity with ultra-sensitive fluorescent probes.
Global analysis of protein tyrosine phosphatase activity with ultra-sensitive fluorescent probes.
复制标题
使用超灵敏荧光探针对蛋白质酪氨酸磷酸酶活性进行整体分析。
DOI：
10.1021/pr050449x
复制
发表时间：
2006
影响因子：
4.4
通讯作者：
Zhang,Zhong-Yin
中科院分区：
文献类型：
--
作者：
Kumar,Sanjai;Zhou,Bo;Liang,Fubo;Yang,Heyi;Wang,Wei-Qing;Zhang,Zhong-Yin
Protein tyrosine phosphatases (PTPs) consist of a large family of enzymes known to play important roles in controlling virtually all aspects of cellular processes. However, assigning functional significance of PTPs in normal physiology and in diseases remains a major challenge in cell signaling. Since the function of a PTP is directly associated with its intrinsic activity, which is subject to post-translational regulation, new tools are needed to monitor the dynamic activities of PTPs, rather than mere abundance, on a global scale within the physiologically relevant environment of cells. To meet this objective, we report the synthesis and characterization of two rhodamine-conjugated probes that covalently label the active site of the PTPs in an activity-dependent manner, thus providing a direct readout of PTP activity and superior sensitivity, robustness, and quantifiability to previously reported biotinylated probes. We present evidence that the fluorescent probes can be used to identify new PTP markers and targets for potential diagnosis and treatment of human diseases. We also show that the fluorescent probes are capable of monitoring H2O2-mediated PTP inactivation, which should facilitate the study of regulated H2O2production as a new tier of control over tyrosine phosphorylation-dependent signal transduction. The ability to profile the entire PTP family on the basis of changes in their activity is expected to yield new functional insights into pathways regulated by PTPs and contribute to the discovery of PTPs as novel therapeutic targets.Keywords: PTP • activity-based probes • activity-based proteomics • fluorescent probes • proteomics • PTP activity profiling
