caa a22 4500 445853832 CHVBK 20180317145422.0 cr unu---uuuuu 170323e20110801xx s 000 0 eng 10.1007/s00204-010-0631-8 doi (NATIONALLICENCE)springer-10.1007/s00204-010-0631-8 Purification and characterization of a cytotoxic neurotoxin-like protein from Naja haje haje venom that induces mitochondrial apoptosis pathway [Elektronische Daten] [Amr El Hakim, Amira Gamal-Eldeen, Yasser Shahein, Nahla Mansour, Ahmed Wahby, Amira Abouelella] This study reported the purification and characterization of a cytotoxic, neurotoxin-like protein derived from the venom of the Egyptian cobra Naja haje haje, Elapidae family, and explored their mechanistic role in the cell death. The protein purification was performed through ion-exchange chromatography and gel-filtration chromatography and was characterized by SDS-PAGE, amino acid sequencing, and mass spectrum analysis. The antitumor activity of Naja haje venom (NHV) and its fractions (NHVI, NHV-Ia, NHV-Ib, NHV-Ic, NHV-II, NHV-III, and NHV-IV) were tested against different human cancer cell lines. The molecular cascade of cell death was explored through evaluation of apoptosis/necrosis ratio, DNA fragmentation, histone deacetylase (HDAC) activity, mitochondrial transmembrane potential (Δψm), cytochrome c release, total caspases, caspase-3, caspase-9, and cell cycle analysis by flow cytometry. Most of the separated fractions possessed variable cytotoxic effect against different cancer cells. The most potent antitumor fraction was NHV-Ic due to its ability to induce DNA damaging and fragmentation that was associated with a significant induction of apoptosis via mitochondrial pathway and disturbed cell cycle phases as well as an inhibition of HDAC activity. NHV-Ic induced the mitochondrial pathway initially by the impairment of Δψm besides the DNA damage and in response to that the mitochondria-released cytochrome c that may in turn activated total caspases, caspase-3 and caspase-9 in lymphoblastic leukemia 1301 cells. The partial amino acid sequencing of NHV-Ic revealed 100, 95.65, and 91.3% homology with the Long neurotoxin 1 from Naja haje anchietae (Angolan cobra), Naja haje haje (Egyptian cobra), and Boulengerina annulata annulata (banded water cobra), respectively. Springer-Verlag, 2011 Naja haje haje nationallicence Venom nationallicence Mitochondrial membrane potential nationallicence Apoptosis nationallicence Cytochrome c nationallicence Histone deacetylases nationallicence Caspases nationallicence Cell cycle nationallicence Flow cytometry nationallicence El Hakim Amr Department of Molecular Biology, National Research Center, Dokki, Cairo, Egypt aut Gamal-Eldeen Amira Cancer Biology Laboratory, Center of Excellence for Advanced Sciences, National Research Center, 12622, Dokki, Cairo, Egypt aut Shahein Yasser Department of Molecular Biology, National Research Center, Dokki, Cairo, Egypt aut Mansour Nahla Gut Microbiology and Immunology Laboratory, Center of Excellence for Advanced Sciences, National Research Center, Dokki, Cairo, Egypt aut Wahby Ahmed Department of Molecular Biology, National Research Center, Dokki, Cairo, Egypt aut Abouelella Amira Radiation Biology Department, National Center for Radiation Research and Technology (NCRRT), Nasr City, Cairo, Egypt aut Archives of Toxicology Springer-Verlag 85/8(2011-08-01), 941-952 0340-5761 85:8<941 2011 85 204 https://doi.org/10.1007/s00204-010-0631-8 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00204-010-0631-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- El Hakim Amr Department of Molecular Biology, National Research Center, Dokki, Cairo, Egypt aut NATIONALLICENCE 700 1- Gamal-Eldeen Amira Cancer Biology Laboratory, Center of Excellence for Advanced Sciences, National Research Center, 12622, Dokki, Cairo, Egypt aut NATIONALLICENCE 700 1- Shahein Yasser Department of Molecular Biology, National Research Center, Dokki, Cairo, Egypt aut NATIONALLICENCE 700 1- Mansour Nahla Gut Microbiology and Immunology Laboratory, Center of Excellence for Advanced Sciences, National Research Center, Dokki, Cairo, Egypt aut NATIONALLICENCE 700 1- Wahby Ahmed Department of Molecular Biology, National Research Center, Dokki, Cairo, Egypt aut NATIONALLICENCE 700 1- Abouelella Amira Radiation Biology Department, National Center for Radiation Research and Technology (NCRRT), Nasr City, Cairo, Egypt aut NATIONALLICENCE 773 0- Archives of Toxicology Springer-Verlag 85/8(2011-08-01), 941-952 0340-5761 85:8<941 2011 85 204 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer