Discovery and validation of PURA as a transcription target of 20(S)-protopanaxadiol: Implications for the treatment of cognitive dysfunction

다국어 초록 (Multilingual Abstract)

Background: 20(S)-protopanaxadiol (PPD), a ginsenoside metabolite, has prominent benefits for thecentral nervous system, especially in improving learning and memory. However, its transcriptionaltargets in brain tissue remain unknown.
Methods: In this study, we first used mass spectrometry-based drug affinity responsive target stability(DARTS) to identify the potential proteins of ginsenosides and intersected them with the transcriptionfactor library. Second, the transcription factor PURA was confirmed as a target of PPD by biolayerinterferometry (BLI) and molecular docking. Next, the effect of PPD on the transcriptional levels of targetgenes of PURA in brain tissues was determined by qRT-PCR. Finally, bioinformatics analysis was used toanalyze the potential biological features of these target proteins.
Results: The results showed three overlapping transcription factors between the proteomics of DARTSand transcription factor library. BLI analysis further showed that PPD had a higher direct interaction withPURA than parent ginsenosides. Subsequently, BLI kinetic analysis, molecular docking, and mutations inkey amino acids of PURA indicated that PPD specifically bound to PURA. The results of qRT-PCR showedthat PPD could increase the transcription levels of PURA target genes in brain. Finally, bioinformaticsanalysis showed that these target proteins were involved in learning and memory function.
Conclusion: The above-mentioned findings indicate that PURA is a transcription target of PPD in brain,and PPD upregulate the transcription levels of target genes related to cognitive dysfunction by bindingPURA, which could provide a chemical and biological basis for the study of treating cognitive impairmentby targeting PURA.