Efficient Screening of nNOS-PSD95 Uncoupling Agents Based on Radiometric Fluorescent Molecularly Imprinted Sensors

Xicheng Yang, Yankun Gao, Hongjuan Zhang, Hongliang Xin

Abstract


Novel and efficient ratiometric fluorescent molecularly imprinted sensors (RFMIS) based on epitopes were developed, which can be used for the sensitive detection of neuronal nitric oxide synthase in the screening of neuronal nitric oxide synthase-postsynaptic density95 (nNOS-PSD95) coupling inhibitors. Under appropriate conditions, the fluorescence of the carbon dots quenched with the increasing concentration of nNOS1-133, while the fluorescence of the quantum dots remained unchanged. The fluorescence ratio had a good linearity in the concentration range of 0-500 ng mL-1 for nNOS1-133 and the determination limit was 0.14 ng mL-1. Using the classical nNOS-PSD95 coupling inhibitor (ZL006) as a control, the RFMIS were used as the detector to detect the free nNOS released by Gnetol and 2,3,5,4ʹ-tetrahydroxystilbene-2-O-β-Dglucoside from natural medicine after inhibition of nNOS-PSD95. The results have shown that the uncoupling efficiencies was consistent with co-immunoprecipitation experiments. The study provides a new idea and a new way for efficient screening of nature nNOS-PSD95 coupling inhibitors from natural medicine with the advantages of high efficiency, sensitivity and traceability.

Keywords


Molecularly Imprinted Polymers; Ratio Fluorescent Sensor; Rapid Screening; Ischemic Stroke

Full Text:

PDF

References


Pei, LS., Shen, X., Yan, YG., et al. The molecular mechanism of stroke treatment by Fufang Longmai Ningfang based on network pharmacology[J]. Acta Pharmaceutica Sinica, 2020, 55(5):898-906.

Landman, TRJ., Schoon, Y., Warle, MC., et al. Remote ischemic conditioning as an additional treatment for acute ischemic stroke: the preclinical and clinical evidence[J]. Stroke,2019,50(7):1934-1939.

Qu, WR., Liu, NK., Wu, XB., et al. Disrupting nNOS-PSD95 interaction improves neurological and cognitive recoveries after traumatic brain injury[J]. Cerebral Cortex, 2020, 30(7): 3859-3871.

Smith, AE., Xu, Z., Lai, YY., et al. Source memory in rats is impaired by an NMDA receptor antagonist but not by PSD95-nNOS protein–protein interaction inhibitors[J]. Behavioural Brain Research, 2016,305:23-29.

Lin, JS., Lai, EM., Protein–protein interactions: Co-Immunoprecipitation. bacterial protein secretion systems[J]. Methods Molecular Biology, 2017, 1615:211-219.

Katz, A., Davis, ME., Molecular imprinting of bulk, microporous silica[J]. Nature, 2000, 403(6767): 286–289.

Pratama, KF., Manik, MER., Rahayu, D., et al. Effect of the molecularly imprinted polymer component ratio on analytical performance[J]. Chemical and Pharmaceutical Bulletin, 2020, 68(11):1013-1024.

S. Patra, R. Choudhary, E. Roy, et al. Triple signalling mode carbon dots-based biodegradable molecularly imprinted polymer as a multitasking visual sensor for rapid and “on-site” monitoring of silver ions[J]. Journal of Materials Chemistry C, 2017,5,11965-11976.

Zhou, ZP., Li, T., Xu, WZ., et al. Synthesis and characterization of fluorescence molecularly imprinted polymers as sensor for highly sensitive detection of dibutyl phthalate from tap water samples[J]. Sensor Actuators B,2017,240, 1114-1122.




DOI: http://dx.doi.org/10.18686/aem.v11i3.295

Refbacks

  • There are currently no refbacks.


Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.