title: Purification of nasulysin-1: A new toxin from Porthidium nasutum snake venom that specifically induces apoptosis in leukemia cell model through caspase-3 and apoptosis-inducing factor activation.
abstract: Nasulysin-1, a new zinc-metalloproteinase from the snake venom of the hognose pit viper Porthidium nasutum, was purified to homogeneity using molecular exclusion chromatography and high performance liquid chromatography on a reverse phase column. The molecular mass of the purified enzyme was 25,900 kDa and pI 4.1, as determined by 1D and 2D polyacrylamide gel electrophoresis. Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) analysis of the N-terminal amino acid sequence (1FSPRYIELVVVADHGMFKKYNSNLNTIR28; 1TASLANLEVWSK12; 1DLLPR6) of the purified nasulysin-1, shows close structural homology with other snake venom metalloproteinases isolated from different snake venoms. The purified nasulysin-1 showed specific apoptosis-inducing activity in Jurkat and K562 cells, a T-cell acute lymphocytic leukemia (ALL) and chronic myeloid leukemia (AML) cell model, respectively, without affecting the viability of human lymphocyte cells. After 48 h treatment, nasulysin-1 (20 μg/mL) induced loss of the mitochondrial membrane potential (ΔΨm), activated the apoptosis-inducing factor (AIF), activated the protease caspase-3, and induced chromatin condensation and DNA fragmentation, all hallmarks of apoptosis. These results strongly suggest that nasulysin-1 selectively induces apoptosis to eliminate leukemia cells. Thus, these data warrant further investigation into the use of the metalloproteinase protein, nasulysin-1 as a potential therapeutic agent for treating leukemia.
authors.affiliation: Neuroscience Research Group, Medical Research Institute, Faculty of Medicine, University of Antioquia, Calle 70 N° 52-21, and Calle 62 #52-59, Building 1, Room 412, SIU, P.O. Box 1226, Medellín, Colombia.
authors.forename: Angelica Rocio
authors.initials: AR
authors.lastname: Bonilla-Porras
authors.affiliation: Ofidism and Scorpionism Program, University of Antioquia, Calle 70 N° 52-21, and Calle 62 #52-59, Building 1, Room 631, SIU, P.O. Box 1226, Medellin, Colombia.
authors.forename: Leidy Johana
authors.initials: LJ
authors.lastname: Vargas
authors.affiliation: Neuroscience Research Group, Medical Research Institute, Faculty of Medicine, University of Antioquia, Calle 70 N° 52-21, and Calle 62 #52-59, Building 1, Room 412, SIU, P.O. Box 1226, Medellín, Colombia.
authors.forename: Marlene
authors.initials: M
authors.lastname: Jimenez-Del-Rio
authors.affiliation: Ofidism and Scorpionism Program, University of Antioquia, Calle 70 N° 52-21, and Calle 62 #52-59, Building 1, Room 631, SIU, P.O. Box 1226, Medellin, Colombia; School of Microbiology, University of Antioquia, Calle 70 N° 52-21, P.O. Box 1226, Medellin, Colombia.
authors.forename: Vitelbina
authors.initials: V
authors.lastname: Nuñez
authors.affiliation: Neuroscience Research Group, Medical Research Institute, Faculty of Medicine, University of Antioquia, Calle 70 N° 52-21, and Calle 62 #52-59, Building 1, Room 412, SIU, P.O. Box 1226, Medellín, Colombia. Electronic address: calberto.velez@udea.edu.co.
authors.forename: Carlos
authors.initials: C
authors.lastname: Velez-Pardo
mesh_terms: D000595:Amino Acid Sequence; D017209:Apoptosis / Q000187:drug effects*; D051033:Apoptosis Inducing Factor / Q000378:metabolism*; D001483:Base Sequence; D053148:Caspase 3 / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D019169:Jurkat Cells; D020014:K562 Cells; D007938:Leukemia / Q000201:enzymology / Q000378:metabolism / Q000473:pathology*; D017386:Sequence Homology, Amino Acid; D021241:Spectrometry, Mass, Electrospray Ionization; D014757:Viper Venoms / Q000737:chemistry*
chemical_list: D051033:Apoptosis Inducing Factor; D014757:Viper Venoms; C000619000:nasulysin-1, Porthidium nasutum; D053148:Caspase 3
keywords: Apoptosis; Jurkat; Leukemia; Porthidium nasutum; Venom; Vipera toxins