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Eruca sativa seed napin structural insights and thorough functional characterization
Binish Khaliq1,2, Sven Falke3, Qamar Saeed4
1Botany Division, Institute of Pure and Applied Biology, Bahauddin Zakariya University, Multan, Pakistan.
Scientific Reports
|December 16, 2021
Summary
Rocket salad napin protein (EsNap) shows potent antifungal, anti-cancer, and insecticidal properties. This study characterizes EsNap, revealing its potential for developing novel biocontrol and therapeutic agents.
Area of Science:
- Plant biochemistry and molecular biology
- Protein characterization and structural analysis
- Biopesticide and pharmaceutical research
Background:
- Napin proteins are storage proteins found in cruciferous plants.
- Rocket salad (Eruca sativa) is a widely cultivated plant with potential bioactive compounds.
- Characterization of plant-derived proteins can reveal novel biological activities.
Purpose of the Study:
- To purify and thoroughly characterize the napin protein from Eruca sativa seeds (EsNap).
- To investigate the structural properties of EsNap using biophysical techniques.
- To evaluate the biological activities of EsNap, including antifungal, cytotoxic, and entomotoxic effects.
Main Methods:
- Protein purification using ammonium sulfate precipitation and size-exclusion chromatography.
- Protein characterization via SDS-PAGE, Nano LC-MS/MS, CD spectroscopy, dynamic light scattering, and Small-Angle X-ray Scattering (SAXS).
- Homology modeling using Brassica napus protein structure.
- Biological activity assays against Fusarium graminearum, Huh7 cell line, and Tribolium castaneum.
Main Results:
- EsNap was purified and identified with a molecular weight of approximately 16 kDa, showing 100% sequence identity to napin-3.
- CD spectroscopy revealed a dominant alpha-helical structure, and SAXS data indicated an elongated monomeric shape.
- EsNap demonstrated significant antifungal activity against Fusarium graminearum, cytotoxicity against Huh7 cells (IC50 = 20.49 µM), and strong entomotoxicity against Tribolium castaneum.
Conclusions:
- EsNap is a potent protein with significant antifungal, anti-cancer, and insect resistance properties.
- The structural and biological data provide a foundation for developing EsNap as a novel agent.
- Eruca sativa seeds are a valuable source for discovering new bioactive proteins for agricultural and medicinal applications.
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