title: Cloning and functional expression of secreted phospholipases A(2) from Bothrops diporus (Yarará Chica).
abstract: Bothrops diporus is a very common viper in Argentina. At present, no complete sequence of secreted phospholipase A(2) (sPLA(2)) from this snake has been reported. We have cloned two sPLA(2) isoenzymes as well as a putative sPLA(2)-like myotoxin from venom gland. The two sPLA(2) were expressed as inclusion bodies in Escherichia coli with an N-terminal tag of ubiquitin. After in vitro renaturation and cleavage step, using an ubiquitin specific peptidase, the recombinants exhibited sPLA(2) activity when analyzed by means of Langmuir dilauroylphosphatidylcholine monolayers as substrate. Both enzymes have a similar surface pressure-activity profile when compared with non-recombinant purified isoforms. To our knowledge, this is the first time that analysis of optimal lateral pressure of substrate monolayers by using the surface barostat technique is performed on recombinant sPLA(2)s.
authors.affiliation: Centro de Investigaciones en Química Biológica de Córdoba (CIQUIBIC, UNC-CONICET), Departamento de Química Biológica, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Haya de la Torre y Medina Allende, Ciudad Universitaria, X5000HUA Córdoba, Argentina.
authors.forename: Pablo Javier
authors.initials: PJ
authors.lastname: Yunes Quartino
authors.forename: José Luis
authors.initials: JL
authors.lastname: Barra
authors.forename: Gerardo Daniel
authors.initials: GD
authors.lastname: Fidelio
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017837:Bothrops / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D006868:Hydrolysis; D008969:Molecular Sequence Data; D054497:Phospholipases A2, Secretory / Q000737:chemistry / Q000235:genetics*; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D054497:Phospholipases A2, Secretory