MRI biosensor for protein kinase A encoded by a single synthetic gene.
MRI biosensor for protein kinase A encoded by a single synthetic gene.
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DOI：
10.1002/mrm.24483
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发表时间：
2012-12
影响因子：
3.3
通讯作者：
Gilad, Assaf A.
中科院分区：
文献类型：
--
作者：
Airan, Raag D.;Bar-Shir, Amnon;Liu, Guanshu;Pelled, Galit;McMahon, Michael T.;van Zijl, Peter C. M.;Bulte, Jeff W. M.;Gilad, Assaf A.
包括蛋白激酶A（PKA）在内的蛋白激酶是无数重要信号通路的基础。在遗传特异性细胞群体中以高空间分辨率在体内和实时监测激酶活性的能力是尚未满足的需求，对于理解复杂的生物系统以及临床前开发和筛选新疗法至关重要。使用的假设，蛋白激酶的天然识别序列可以使用化学交换饱和转移（CEST）磁共振成像（MRI）检测，我们设计了一个基因编码的生物传感器组成的8个串联重复的肽LRRASLG，PKA的天然目标。该传感器显示了PKA磷酸化后CEST信号的可测量变化。天然PKA底物LRRASLG在+1.8和+3.6 ppm处表现出CEST-MRI对比度，磷酸化后变化>50%，时间分辨率为分钟级。编码LRRASLG的八个单体的合成基因的表达在这些CEST频率处产生两个峰。总之，这些结果表明，该基因可用于测定PKA水平在生物学相关系统。重要的是，用于该特定传感器的设计策略可以适用于临床上感兴趣的蛋白激酶的宿主。
Protein kinases including Protein Kinase A (PKA) underlie myriad important signaling pathways. The ability to monitor kinase activity in vivo and in real-time with high spatial resolution in genetically-specified cellular populations is a yet unmet need, crucial for understanding complex biological systems as well as for preclinical development and screening of novel therapeutics. Using the hypothesis that the natural recognition sequences of protein kinases may be detected using chemical exchange saturation transfer (CEST) magnetic resonance imaging (MRI), we designed a genetically encoded biosensor composed of eight tandem repeats of the peptide LRRASLG, a natural target of PKA. This sensor displays a measurable change in CEST signal following phosphorylation by PKA. The natural PKA substrate LRRASLG exhibits a CEST-MRI contrast at +1.8 and +3.6 ppm, with a >50% change after phosphorylation with minutes-scale temporal resolution. Expression of a synthetic gene encoding eight monomers of LRRASLG yielded two peaks at these CEST frequencies. Taken together, these results suggest that this gene may be used to assay PKA levels in a biologically relevant system. Importantly, the design strategy used for this specific sensor may be adapted for a host of clinically interesting protein kinases.
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影响因子：
3.3
作者：
McMahon, Michael T.;Gilad, Assaf A.;DeLiso, Marco A.;Berman, Stacey D. Cromer;Bulte, Jeff W. M.;van Zijl, Peter C. M.
通讯作者：
van Zijl, Peter C. M.
影响因子：
3.3
作者：
Jones, Craig K.;Polders, Daniel;Hua, Jun;Zhu, He;Hoogduin, Hans J.;Zhou, Jinyuan;Luijten, Peter;van Zijl, Peter C. M.
通讯作者：
van Zijl, Peter C. M.
影响因子：
3.3
作者：
Singh, Anup;Cai, Kejia;Haris, Mohammad;Hariharan, Hari;Reddy, Ravinder
通讯作者：
Reddy, Ravinder
影响因子：
15
作者：
Liu, Guanshu;Liang, Yajie;Bar-Shir, Amnon;Chan, Kannie W. Y.;Galpoththawela, Chulani S.;Bernard, Segun M.;Tse, Terence;Yadav, Nirbhay N.;Walczak, Piotr;McMahon, Michael T.;Bulte, Jeff W. M.;van Zijl, Peter C. M.;Gilad, Assaf A.
通讯作者：
Gilad, Assaf A.
通过CA2+-CAMP-PKA振荡电路实现的PKA的信号多样性。
影响因子：
14.8
作者：
Ni, Qiang;Ganesan, Ambhighainath;Aye-Han, Nwe-Nwe;Gao, Xinxin;Allen, Michael D.;Levchenko, Andre;Zhang, Jin
通讯作者：
Zhang, Jin