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Establishment of a screening model for compounds uncoupling the interaction of nNOS with PSD-95
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Lei ZHANG, He-ming ZHOU, Xiao-yun LI, Chu ZHANG, Ya-zhuo ZHANG, Miao ZHOU, Li-na CHEN, Yu ZHANG*
Acta Pharmaceutica Sinica | 2019, 54(4) : 638 - 644
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Acta Pharmaceutica Sinica | 2019, 54(4): 638-644
• Original Articles •
Establishment of a screening model for compounds uncoupling the interaction of nNOS with PSD-95
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Lei ZHANG, He-ming ZHOU, Xiao-yun LI, Chu ZHANG, Ya-zhuo ZHANG, Miao ZHOU, Li-na CHEN, Yu ZHANG*
Affiliations
  • School of Pharmacy, Nanjing Medical University, Nanjing 211166, China
Published: 2019-04-12 doi: 10.16438/j.0513-4870.2018-0935
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In ischemic stroke, increased level of neuronal complex of nitric oxide synthase (nNOS)-postsynaptic density protein-95 (PSD-95) plays an important role in neuronal damage. We aimed to establish a screening model to identify compounds capable of uncoupling nNOS interaction with PSD-95. In this model, human embryonic kidney-293T (HEK-293T) cells were transfected with either pCDH-Flag-nNOS or pcDNA3.1-PSD-95 plasmid to obtain the protein of Flag-nNOS or PSD-95. Incubating Flag-nNOS with PSD-95 causes formation of the nNOS-PSD-95 complex. ZL006, a known uncoupler of nNOS-PSD-95 interaction, can disturb the interaction between Flag-nNOS and PSD-95, serving as a positive control. The method coupling antibodies to magnetic beads with glutaraldehyde was used to decrease the cost and increase the efficiency. To establish that our model is suitable for selecting nNOS-PSD-95 uncouplers, we evaluated the ability of IC87201, another reported uncoupler of nNOS-PSD-95 interaction, and structural analogs of ZL006. IC87201 and one structure analog of ZL006 showed uncoupling effect, supporting that our model can be used to select different types uncoupler blocking nNOS-PSD-95 interaction.

nNOS-PSD-95  /  uncoupler  /  ZL006  /  screening model
Lei ZHANG, He-ming ZHOU, Xiao-yun LI, Chu ZHANG, Ya-zhuo ZHANG, Miao ZHOU, Li-na CHEN, Yu ZHANG. Establishment of a screening model for compounds uncoupling the interaction of nNOS with PSD-95[J]. Acta Pharmaceutica Sinica, 2019 , 54 (4) : 638 -644 . DOI: 10.16438/j.0513-4870.2018-0935
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doi: 10.16438/j.0513-4870.2018-0935
  • Receive Date：2018-10-16
  • Online Date：2026-01-26
  • Published：2019-04-12
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  • Received：2018-10-16
  • Revised：2018-11-17
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    School of Pharmacy, Nanjing Medical University, Nanjing 211166, China
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小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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