Abstract
Tetradecyl 2,3-dihydroxybenzoate (ABG-001) is a lead compound derived from gentisides with a remarkable neuritogenic activity. However, the target of ABG-001 is yet to be defined to date. In this study, the potential target of ABG-001 was investigated via an activity-based protein profiling (ABPP) analysis, which is a chemical proteomic method for target identification by using chemical probes. Results indicated that the potential target proteins of ABG-001 were heat shock cognate 70 kDa protein (Hsc70), 78 kDa glucose-regulated protein (GRP78), and 14-3-3 theta protein. Then, the potential target of ABG-001 was confirmed by using inhibitors, the cellular thermal shift assay (CETSA) and small-interfering RNA (siRNA) analysis. The inhibitor of Hsc70 and siRNA significantly decreased the neurite outgrowth induced by ABG-001. Furthermore, ABG-001 induced neurite outgrowth was reduced by siRNA against Hsc70, and the results of CETSA suggested that Hsc70 showed a significant thermal stability-shifted effect upon ABG-001 treatment. These results indicated that Hsc70 is the target protein of ABG-001 in PC12 cells.
Highlights
Neuritogenic Effect in PC12 Cells.Nerve growth factor (NGF), one of the most significant neurotrophic factors that vitally contributes to neuronal growth, survival, differentiation, and function maintenance, was once considered as an ideal therapeutic agent for the treatment of neurodegenerative diseases, such as Alzheimer’s disease (AD) [1]
P4htm related to metal ion binding, Fxyd6 related to ion transport, Hspa8 related to neuron protection, Napa and Akt related to neuron differentiation were significantly down-regulated after being treated with ABG-001
PC12 cells were treated with negative control, ABG-P1, and ABG-PC
Summary
ABG-001, a biologically active small molecule with excellent neuritogenic activity similar to NGF on PC12 cells, has significant potential for drug development to treat neurodegenerative diseases. Despite that ideal phenotypic screening results were obtained, the unknown mechanism of action study and target identity of the neuritogenic activity induced by ABG-001 still remained to be investigated, which are significantly crucial due to their correlations with the development of small-molecule probes and their application in biology and drug discovery [6]. The probes, designed according to the structure of the active leading compound, would bind to and covalently react with its target protein via an electrophilic trap or a photocrosslinking group in the living cells. The results indicated that ABG-001 potentially targeted the Hsc protein
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
