PNAS ASSAY：科学家鉴别出记忆障碍的新型治疗靶点

作者：张馨予

日前，发表在国际杂志PNAS及ASSAY and Drug Development Technologies上的两篇研究报道中，来自斯克里普斯研究所的研究人员鉴别出了一系列记忆障碍的新型治疗靶点，同时开发出了新型的筛选测试技术有望筛选出抵御记忆障碍的新型化合物，相关研究或为开发治疗老化相关疾病的新型疗法带来希望，比如阿尔兹海默氏症和痴呆症等。

2014年11月14日讯 /生物谷BIOON/ --日前，发表在国际杂志PNAS及ASSAY and Drug Development Technologies上的两篇研究报道中，来自斯克里普斯研究所的研究人员鉴别出了一系列记忆障碍的新型治疗靶点，同时开发出了新型的筛选测试技术有望筛选出抵御记忆障碍的新型化合物，相关研究或为开发治疗老化相关疾病的新型疗法带来希望，比如阿尔兹海默氏症和痴呆症等。

生物学家Sathyanarayanan V. Puthanveettil说道，目前我们正在寻找对记忆形成很关键的分子，在上述两篇研究报道中，我们发现了新型的路径可以帮助我们鉴别新型候选药物来治疗记忆障碍等相关疾病。

揭秘突触蛋白组

发表在PNAS的研究论文中，研究者详细描述了一系列可以运输至突触的蛋白质，这些蛋白质组名为突触蛋白质组；突触是神经元的一部分，在工作期间（比如记忆存储）其可以将电化学信号传送至其它细胞中，而本文研究中揭示了突触的工作机制，以及其组成如何随着学习而变化，随后研究人员还揭示了大脑疾病如何影响突触的正常工作。

研究者表示，我们重点对驱动蛋白进行研究，其对于细胞特殊蛋白的运输至关重要，通过对三种驱动蛋白结构的分析，研究人员发现，大约40%至50%的蛋白质“货物”都是突触蛋白质，而驱动蛋白的识别也可以帮助决定被运输的蛋白质类型，研究结果揭示了驱动蛋白在调节整个突触蛋白组组成上的重要作用。

研究者Xin-An Liu表示，我们首次研究揭示了相同神经元中驱动蛋白的表达如何携带大量不同的蛋白质“货物”，利用该机制我们就可以鉴别出记忆等主要障碍的的靶点分子，下一步我们将继续研究来解释突触蛋白质组在神经疾病发生过程中的改变机制。

新型药物候选者

发表在ASSAY上的研究论文中，研究人员描述了一种高通量的筛选检测技术，其可以基于记忆障碍疗法中的驱动蛋白和轴突输送来筛选潜在的治疗药物；Puthanveetil博士表示，我们开发的荧光检测实验具有强大和高效的可重复性，基于此我们就可以筛选鉴别出治疗人类记忆障碍的新型分子药物。（生物谷Bioon.com）

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doi:10.1073/pnas.1401483111

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New approach to capture and characterize synaptic proteome

Xin-An Liua, Beena Kadakkuzhaa,1, Bruce Pascalb,1, Caitlin Stecklerc, Komolitdin Akhmedova, Long Yand, Michael Chalmersc, and Sathyanarayanan V. Puthanveettila,2

Little is known regarding the identity of the population of proteins that are transported and localized to synapses. Here we describe a new approach that involves the isolation and systematic proteomic characterization of molecular motor kinesins to identify the populations of proteins transported to synapses. We used this approach to identify and compare proteins transported to synapses by kinesin (Kif) complexes Kif5C and Kif3A in the mouse hippocampus and prefrontal cortex. Approximately 40–50% of the protein cargos identified in our proteomics analysis of kinesin complexes are known synaptic proteins. We also found that the identity of kinesins and where they are expressed determine what proteins they transport. Our results reveal a previously unappreciated role of kinesins in regulating the composition of synaptic proteome.

doi:10.1089/adt.2014.579

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High-Throughput Screening for Small Molecule Modulators of Motor Protein Kinesin

Kadakkuzha Beena M., Spicer Timothy P., Chase Peter, Richman Jeffery B., Hodder Peter, and Puthanveettil Sathyanarayanan V

The kinesin superfamily of motor proteins are involved in the active transport of a large number of cargos such as organelles, proteins, and RNAs from the neuronal cell body to distal neuronal processes. Previously, we have shown that kinesin-mediated axonal transport of proteins and RNAs are important for long-term memory storage. Identification of small molecules that can activate or inhibit kinesins is of specific interest due to the significance of kinesin-mediated functions in neuronal health and plasticity. Here, we describe a high-throughput screening assay designed to specifically identify compounds that inhibit or activate adenosine triphosphatase activity of the kinesin 5B of humans. The luminescence-based assay that we developed is highly reproducible and robust. Using this approach, we screened a pharmacologically characterized compound collection and have identified small molecules with either activator or inhibitor-like activity. To further characterize screening hits, we also developed an orthogonal assay based on absorbance and a counter screen assay based on luminescence. Development of such assays is important to help identify small molecules that can be used in potential drug development efforts targeted at modulating the function of kinesin.