Rapid, Direct SMOOTHENED Activity Assays in Live Cells Using cAMP-Based Conformational Sensors.
Rapid, Direct SMOOTHENED Activity Assays in Live Cells Using cAMP-Based Conformational Sensors.
复制标题
使用基于 cAMP 的构象传感器对活细胞进行快速、直接的平滑活性测定。
DOI：
10.1007/978-1-0716-1701-4_15
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Myers,BenjaminR
中科院分区：
文献类型：
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作者：
Walker,MadisonF;Myers,BenjaminR
PATCHED 1（PTCH 1）和SMOOTHENED（SMO）之间的通讯是发育和疾病中Hedgehog（Hh）信号转导的基础。我们描述了一种基于SMO活性依赖性细胞cAMP浓度变化的实时细胞SMO功能测定。该测定能够在Hh配体灭活PTCH 1的几分钟内检测SMO构象的变化。因此，它扩大了可用于剖析PTCH 1-SMO通信的实验扰动范围，从而能够更深入地了解Hh信号转导中长期存在的奥秘。
Communication between PATCHED1 (PTCH1) and SMOOTHENED (SMO) is fundamental to Hedgehog (Hh) signal transduction in development and disease. We describe a real-time cell-based SMO functional assay based on SMO activity-dependent changes in cellular cAMP concentrations. This assay is capable of detecting changes in SMO conformation within minutes of PTCH1 inactivation by Hh ligands. As a result, it expands the range of experimental perturbations that can be used to dissect PTCH1-SMO communication, enabling a deeper mechanistic understanding of a longstanding mystery in Hh signal transduction.