title: Comprehensive analysis of venom from the scorpion Centruroides tecomanus reveals compounds with antimicrobial, cytotoxic, and insecticidal activities.
abstract: Centruroides tecomanus is a medically important scorpion of the state of Colima (Mexico). This communication reports the identification of venom components of this scorpion with biological activity over insects/crickets (Acheta domestica), crustaceans/fresh water shrimps (Cambarellus montezumae), and mammalians/mice (Mus musculus, strain CD1). It also describes the pharmacological effects on cell lines in culture (L5178Y cells, HeLa cells, HuTu cells and Jurkat E6-1 cells), as well as on several types of bacteria (see below). The soluble venom of this scorpion was fractionated by high-performance liquid chromatography (HPLC) and collected separately in twelve independent fractions collected over 60 min run (5 min time apart each other). The HPLC components of fraction VII were lethal to all three species used for assay. The IVth fraction had a toxic effect on freshwater shrimps. In this species, fractions VI, VII and VIII were all lethal. For crickets, fractions V and VI were toxic and fraction VII was lethal. In mouse, the lethal components were found in fraction VII, whereas fraction VIII was toxic, but not lethal, at the doses assayed. The molecular weight of peptides from the various group of fractions were identified by mass spectrometry determination. Components lethal to mice showed molecular weights from 7013 to 7487 Da. Two peptides were obtained in homogeneous form and shown to be lethal to the three species of animal used for assay. The soluble venom tested on L5178Y cell line survival was shown to be cytotoxic, at 10-100 μg/mL concentration, when compared to control murine splenocytes (p = 0.007). The soluble venom applied to Hela, Hutu and Jurkat cell lines did not show cytotoxic effects at these concentrations. On the contrary, it seems to have a proliferative effect. However the HPLC fractions I, III, VI and XII do have a cytotoxic effect on Jurkat E06-1 cells in culture at 200 μg/mL concentration. The antimicrobial activity of the venom fractions on Staphylococcus aureus (gram-positive), Escherichia coli, Pseudomonas aeruginosa y Salmonella spp (gram-negative) was measured, using the liquid inhibition growth system. The four strains of bacteria used were susceptible to fractions III and IV, affecting all four bacterial strains at concentrations below 5 μg/mL.
authors.affiliation: Facultad de Ciencias Químicas, Universidad de Colima, Kilometro 9 Carretera Colima-Coquimatlan, Código Postal 28400, Coquimatlan, Colima, Mexico; Facultad de Medicina, Universidad de Colima, Av. Universidad 333, Colonia las Viboras, Código Postal 28040, Colima, Mexico. Electronic address: lauravaldez@ucol.mx.
authors.forename: L L
authors.initials: LL
authors.lastname: Valdez-Velazquéz
authors.affiliation: Facultad de Medicina, Universidad de Colima, Av. Universidad 333, Colonia las Viboras, Código Postal 28040, Colima, Mexico.
authors.forename: M T
authors.initials: MT
authors.lastname: Romero-Gutierrez
authors.affiliation: Facultad de Ciencias Químicas, Universidad de Colima, Kilometro 9 Carretera Colima-Coquimatlan, Código Postal 28400, Coquimatlan, Colima, Mexico; Facultad de Medicina, Universidad de Colima, Av. Universidad 333, Colonia las Viboras, Código Postal 28040, Colima, Mexico.
authors.forename: I
authors.initials: I
authors.lastname: Delgado-Enciso
authors.affiliation: Centro Universitario de Investigaciones Biomédicas, Universidad de Colima, Av. 25 de Julio 965, Código Postal 28045, Colima, Mexico.
authors.forename: O
authors.initials: O
authors.lastname: Dobrovinskaya
authors.affiliation: Facultad de Medicina, Universidad de Colima, Av. Universidad 333, Colonia las Viboras, Código Postal 28040, Colima, Mexico.
authors.forename: V
authors.initials: V
authors.lastname: Melnikov
authors.affiliation: CONACYT Research Fellow - Laboratorio de Ecología Molecular Microbiana, Centro de Investigaciones en Ciencias Microbiológicas-Instituto de Ciencias, Benemérita Universidad Autónoma de Puebla, Ciudad Universitaria, Código Postal 72570, Puebla, Mexico.
authors.forename: V
authors.initials: V
authors.lastname: Quintero-Hernández
authors.affiliation: Facultad de Ciencias, Universidad de Colima, c/Bernal Díaz del Castillo 340, Código Postal 28045, Colima, Mexico.
authors.forename: S G
authors.initials: SG
authors.lastname: Ceballos-Magaña
authors.affiliation: Facultad de Ciencias Químicas, Universidad de Colima, Kilometro 9 Carretera Colima-Coquimatlan, Código Postal 28400, Coquimatlan, Colima, Mexico.
authors.forename: M A
authors.initials: MA
authors.lastname: Gaitan-Hinojosa
authors.affiliation: Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Colonia Chamilpa, Apartado Postal 510-3, Cuernavaca, Morelos, Mexico.
authors.forename: F I
authors.initials: FI
authors.lastname: Coronas
authors.affiliation: Universidad de Guadalajara, Centro Universitario de Ciencias Exactas, Blvd. Gral. Marcelino García Barragán 1421, Olímpica, Apartado Postal 44430, Guadalajara, Jalisco, Mexico.
authors.forename: A M
authors.initials: AM
authors.lastname: Puebla-Perez
authors.affiliation: Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Colonia Chamilpa, Apartado Postal 510-3, Cuernavaca, Morelos, Mexico.
authors.forename: F
authors.initials: F
authors.lastname: Zamudio
authors.affiliation: Facultad de Ciencias Químicas, Universidad de Colima, Kilometro 9 Carretera Colima-Coquimatlan, Código Postal 28400, Coquimatlan, Colima, Mexico.
authors.forename: I
authors.initials: I
authors.lastname: De la Cruz-García
authors.affiliation: Facultad de Ciencias Químicas, Universidad de Colima, Kilometro 9 Carretera Colima-Coquimatlan, Código Postal 28400, Coquimatlan, Colima, Mexico.
authors.forename: O F
authors.initials: OF
authors.lastname: Vázquez-Vuelvas
authors.affiliation: Facultad de Medicina, Universidad de Colima, Av. Universidad 333, Colonia las Viboras, Código Postal 28040, Colima, Mexico.
authors.forename: A D
authors.initials: AD
authors.lastname: Soriano-Hernandez
authors.affiliation: Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Colonia Chamilpa, Apartado Postal 510-3, Cuernavaca, Morelos, Mexico.
authors.forename: L D
authors.initials: LD
authors.lastname: Possani
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000009:adverse effects / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000009:adverse effects / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D060829:Arthropod Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology / Q000633:toxicity; D003400:Astacoidea / Q000187:drug effects / Q000254:growth & development; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D055808:Drug Discovery; D006090:Gram-Negative Bacteria / Q000187:drug effects / Q000254:growth & development; D006135:Gryllidae; D006801:Humans; D007274:Injections, Intraperitoneal; D007306:Insecticides / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology / Q000633:toxicity; D008800:Mexico; D051379:Mice; D008826:Microbial Sensitivity Tests; D012604:Scorpion Venoms / Q000008:administration & dosage / Q000737:chemistry* / Q000633:toxicity; D012605:Scorpions / Q000254:growth & development; D013154:Spleen / Q000166:cytology / Q000187:drug effects; D013211:Staphylococcus aureus / Q000187:drug effects / Q000254:growth & development
chemical_list: D000900:Anti-Bacterial Agents; D000970:Antineoplastic Agents; D060829:Arthropod Proteins; D007306:Insecticides; D012604:Scorpion Venoms
keywords: Antimicrobial activity; Cancer cell; Scorpion venom; Toxin