Purification, Identification and Neuroprotective Effects of Proteins from Bombyx batryticatus in Glu-Stimulated PC12 Cells

Bombyx batryticatus (BB) is one of the most commonly used Traditional Chinese Medicines (TCMs) in treatment convulsions and epilepsy. The antiepileptic effects total proteins from BB (BBPs) have been proven our previous research. In this study, BBPs were further purified, neuroprotective evaluated Glu-stimulated PC12 cells, structure was identified by Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Six subfractions (PF-1 to PF-6) obtained DEAE-52 Sepharose FF ion-exchange chromatography. It found that PF-1, PF-2, PF-3, with similar protein compositions, possessed cells significantly increasing GABA level, decreasing levels IL-1β TNF-α. active fraction (PF-2) separated Sephadex G-75 gel filtration chromatography, an effective component named PF-2-2 obtained. Fluorescein isothiocyanate-labeled (FITC-PF-2-2) prepared, binding FITC-F-2-2 directly observed a confocal microscope. first bind surface then internalize into cells. main band analyzed MALDI-TOF/TOF-MS searched MASCOT database; finally Low molecular 30 kDa lipoprotein 21G1 identified. conclusion, purified isolated potential application prospects