Using "On/Off" F-19 NMR/Magnetic Resonance Imaging Signals to Sense Tyrosine Kinase/Phosphatase Activity in Vitro and in Cell Lysates
Using "On/Off" F-19 NMR/Magnetic Resonance Imaging Signals to Sense Tyrosine Kinase/Phosphatase Activity in Vitro and in Cell Lysates
复制标题
使用“开/关”F-19 NMR/磁共振成像信号检测体外和细胞裂解物中的酪氨酸激酶/磷酸酶活性
DOI：
10.1021/acs.analchem.6b00036
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发表时间：
2016
影响因子：
7.4
通讯作者：
Liang Gaolin
中科院分区：
文献类型：
--
作者：
Zheng Zhen;Sun Hongbin;Hu Chen;Li Gongyu;Liu Xiaomei;Chen Peiyao;Cui Yusi;Liu Jing;Wang Junfeng;Liang Gaolin
酪氨酸激酶和磷酸酶是生物体内两种重要的拮抗酶。发展非侵入性的方法来感测他们的活动具有高的空间和时间分辨率仍然具有挑战性。在此，我们合理地设计了水凝胶因子Nap-Phe-Phe（CF 3）-Glu-Tyr-Ile-OH（1a），其超分子水凝胶（即，凝胶1a）可在酪氨酸激酶的作用下发生解聚，其磷酸盐前体Nap-Phe-Phe（CF 3）-Glu-Tyr（H2 PO 3）-Ile-OH（1b）可在碱性磷酸酶（ALP）的作用下发生自组装，形成超分子水凝胶凝胶Gel 1b。用流变学和低温透射电子显微镜（cryo-TEM）对水凝胶的力学性能和内部纤维网络进行了表征。首次利用具有“On/Off“~（19）FNMR/MRI信号的超分子水凝胶的自组装/自组装技术，在体外和细胞裂解液中检测了这两种重要酶的活性。我们期望我们的新的19 F NMR/磁共振成像（MRI）方法将有助于药物研究人员筛选这两种酶的新抑制剂，而没有空间位阻。
Tyrosine kinase and phosphatase are two important, antagonistic enzymes in organisms. Development of noninvasive approach for sensing their activity with high spatial and temporal resolution remains challenging. Herein, we rationally designed a hydrogelator Nap-Phe-Phe(CF3)-Glu-Tyr-Ile-OH (1a) whose supramolecular hydrogel (i.e., Gel1a) can be subjected to tyrosine kinase-directed disassembly, and its phosphate precursor Nap-Phe-Phe(CF3)-Glu-Tyr(H2PO3)-Ile-OH (1b), which can be subjected to alkaline phosphatase (ALP)-instructed self-assembly to form supramolecular hydrogel Gel1b, respectively. Mechanic properties and internal fibrous networks of the hydrogels were characterized with rheology and cryo transmission electron microscopy (cryo-TEM). Disassembly/self-assembly of their corresponding supramolecular hydrogels conferring respective “On/Off”19F NMR/MRI signals were employed to sense the activity of these two important enzymesin vitroand in cell lysates for the first time. We anticipate that our new19F NMR/magnetic resonance imaging (MRI) method would facilitate pharmaceutical researchers to screen new inhibitors for these two enzymes without steric hindrance.