Purification and characterization of a novel antitumor protein with antioxidant and deoxyribonuclease activity from edible mushroom Pholiota nameko

摘要: A novel antitumor protein from the edible mushroom Pholiota nameko (PNAP) was purified through a two-step chromatographic procedure including SP cation exchange chromatogram and Superdex gel filtration. The approximate molecular weight demonstrated to be 18.5 kDa by SDS-PAGE ultracentrifugation analysis N-terminal sequence detected as AGRTFIGYNG Edman degradation. Biochemical characterization showed that it exhibited significant antioxidant activity effectively scavenging hydroxyl 1,1-diphenyl-2-picrylhydrazyl radicals compared standard butylated hydroxy anisole. PNAP had deoxyribonuclease with optimum pH temperature were 5.0 60 °C respectively, well can act on both double-stranded single-stranded DNA, but preferentially DNA. displayed against cancer cell lines such MCF7 Hela cells. Human breast cells treated produced typical apoptotic morphological changes chromatin condensation, accumulation of sub-G1 alternation mitochondrial permeability. induced apoptosis entailed loss membrane potential resulting in release cytochrome c into cytosol, activation caspase-9 caspase-3, which are responsible for execution apoptosis, implying intrinsic signal pathway is involved apoptosis.