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Identification and characterization of Scp15, a protein from Streptomyces coelicolor A3(2) inducing neurites in PC12
Satoru Nakashima1, Shuji Wakatsuki, Tatsuya Yokoyama
1Department of Biotechnology, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Abstract:
We previously showed that a fungal protein, p15, induces neurite outgrowth and differentiation of rat pheochromocytoma PC12 cells. We report here the identification and characterization of a protein similar to p15, found in Streptomyces coelicolor A3(2). This hypothetical protein, tentatively named Scp15, has significant similarity with p15, including conserved positions of four cysteine residues involved in the formation of essential disulfide bonds in p15. Hexahistidine-tagged recombinant Scp15 proteins were produced in Escherichia coli, purified, and analyzed for their neurite-inducing activity. Although they were less active than p15, they dose-dependently induced neurites and the expression of neurofilament M. Neurite outgrowth by Scp15 was inhibited by nicardipine, suggesting that Scp15 induces neurites via activation of a calcium signaling pathway.
Insights
A novel bacterial protein, Scp15, similar to fungal p15, promotes neurite outgrowth in neuronal cells. This suggests a conserved mechanism for neuronal differentiation involving calcium signaling pathways.
Area of Science:
- Neuroscience
- Microbiology
- Molecular Biology
Background:
- Fungal protein p15 was previously shown to induce neurite outgrowth and differentiation in rat pheochromocytoma PC12 cells.
- Neuronal differentiation is a critical process in nervous system development and function.
Purpose of the Study:
- To identify and characterize proteins similar to fungal p15 in other organisms.
- To investigate the neurite-inducing activity of a newly identified protein, Scp15, from Streptomyces coelicolor A3(2).
Main Methods:
- Sequence analysis to identify proteins with similarity to p15.
- Production and purification of hexahistidine-tagged recombinant Scp15 in E. coli.
- Assay of Scp15's neurite-inducing activity on PC12 cells and analysis of neurofilament M expression.
Main Results:
- Scp15 shares significant similarity with p15, including conserved cysteine residues crucial for disulfide bond formation.
- Recombinant Scp15 dose-dependently induced neurite outgrowth and neurofilament M expression in PC12 cells, albeit with lower activity than p15.
- Nicardipine inhibited Scp15-induced neurite outgrowth, indicating involvement of calcium signaling.
Conclusions:
- Scp15 is a novel bacterial protein with neurite-inducing capabilities.
- Scp15 likely induces neuronal differentiation through the activation of calcium signaling pathways.
- These findings suggest conserved molecular mechanisms for neuronal differentiation across different species.
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