Eruca sativa seed napin structural insights and thorough functional characterization

Abstract

A potent napin protein has been thoroughly characterized from seeds of rocket salad (Eruca sativa). Eruca sativa napin (EsNap) was purified by ammonium sulfate precipitation (70%) and size-exclusion chromatography. Single intact 16 kDa EsNap band was reduced to 11 and 5 kDa bands respectively on SDS-PAGE. Nano LC–MS/MS yielded two fragments comprising of 26 residues which showed 100% sequence identity with napin-3 of Brassica napus. CD spectroscopy indicated a dominant α-helical structure of EsNap. Monodispersity of EsNap was verified by dynamic light scattering, which also confirmed the monomeric status with a corresponding hydrodynamic radius of 2.4 ± 0.2 nm. An elongated ab initio shape of EsNap was calculated based on SAXS data, with an Rg of 1.96 ± 0.1 nm. The ab initio model calculated by DAMMIF with P1 symmetry and a volume of approx. 31,100 nm3, which corresponded to a molecular weight of approximately 15.5 kDa. The comparison of the SAXS and ab initio modeling showed a minimized χ2-value of 1.87, confirming a similar molecular structure. A homology model was predicted using the coordinate information of Brassica napus rproBnIb (PDB ID: 1SM7). EsNap exhibited strong antifungal activity by significantly inhibiting the growth of Fusarium graminearum. EsNap also showed cytotoxicity against the hepatic cell line Huh7 and the obtained IC50 value was 20.49 µM. Further, strong entomotoxic activity was experienced against different life stages of stored grain insect pest T. castaneum. The result of this study shows insights that can be used in developing potential antifungal, anti-cancerous and insect resistance agents in the future using EsNap from E. sativa.

Preparation and characterization of cetuximab-loaded egg serum albumin nanoparticles and their uses as a drug delivery system against Caco-2 colon cancer cells

“…A vertical electrophoresis system (MiniVE, Hoefer, U.S.A.) was used at a 75-V constant voltage for 30 min, then 115 V for 1.5 h in electrophoresis buffer (0.25 mM Tris, 1.92 mM glycine, 1% SDS, and pH = 8.3). The gel was stained with Coomassie brilliant blue buffer for 1 h and washed with a destaining agent (40% methanol, 7% acetic acid, and 53% water) (Khaliq et al 2021).…”

Molecular Characteristics Study of The Plants of Two Species Form The Genus Eruca Mill of The Brassicaceae Family, Which are Growing in Iraq

“…It has been observed that E. sativa attracts a large number of insect pollinators, and this may be attributed to the difference in the amount of nectar and its chemical properties, as well as the number of flowers and pollen grains [16]. In the future, possible antifungal, cancer, and insect resistance medicines may be developed using EsNap of the E. sativa species [17].…”

Seed Priming of Wheat Through Salicylic Acid to Induce Salt Tolerance

“…Under abiotic stress conditions the wheat is highly affected which caused reduction in the yield and productivity. The salt stress caused cell membrane damage, production of toxic chemicals and accumulation of reactive oxygen species in the plant body (Ali et al, 2017;Ali and Malik, 2021;Khaliq et al, 2021;Mustafa et al, 2018;Sarwar et al, 2021). The salt or NaCl stress caused damage in the cell membranes, the use of salicylic acid induce the tolerance in the plant body which may help crop plants to withstand under salt and abiotic stress condition (Aaliya et al, 2016;Ali et al, 2015;Iqra et al, 2020;Majid et al, 2017;Mazhar et al, 2020;Sarwar et al, 2022;Zubair et al, 2016).…”