title: Characterization of the single-stranded DNA binding protein pV(VGJΦ) of VGJΦ phage from Vibrio cholerae.
abstract: pV(VGJΦ), a single-stranded DNA binding protein of the vibriophage VGJΦ was subject to biochemical analysis. Here, we show that this protein has a general affinity for single-stranded DNA (ssDNA) as documented by Electrophoretic Mobility Shift Assay (EMSA). The apparent molecular weight of the monomer is about 12.7kDa as measured by HPLC-SEC. Moreover, isoelectrofocusing showed an isoelectric point for pV(VGJΦ) of 6.82 pH units. Size exclusion chromatography in 150mM NaCl, 50mM sodium phosphate buffer, pH 7.0 revealed a major protein species of 27.0kDa, suggesting homodimeric protein architecture. Furthermore, pV(VGJΦ) binds ssDNA at extreme temperatures and the complex was stable after extended incubation times. Upon frozen storage at -20°C for a year the protein retained its integrity, biological activity and oligomericity. On the other hand, bioinformatics analysis predicted that pV(VGJΦ) protein has a disordered C-terminal, which might be involved in its functional activity. All the aforementioned features make pV(VGJΦ) interesting for biotechnological applications.
authors.affiliation: Department of Molecular Biology, National Center for Scientific Research, Havana, Cuba. alina.falero@cnic.edu
authors.forename: Alina
authors.initials: A
authors.lastname: Falero
authors.forename: Andy
authors.initials: A
authors.lastname: Caballero
authors.forename: Sonia
authors.initials: S
authors.lastname: Trigueros
authors.forename: Celso
authors.initials: C
authors.lastname: Pérez
authors.forename: Javier
authors.initials: J
authors.lastname: Campos
authors.forename: Karen
authors.initials: K
authors.lastname: Marrero
authors.forename: Rafael
authors.initials: R
authors.lastname: Fando
mesh_terms: D001435:Bacteriophages / Q000737:chemistry*; D019295:Computational Biology; D004268:DNA-Binding Proteins / Q000737:chemistry*; D007526:Isoelectric Point; D008970:Molecular Weight; D055503:Protein Multimerization; D014734:Vibrio cholerae / Q000737:chemistry*; D014764:Viral Proteins / Q000737:chemistry*
chemical_list: D004268:DNA-Binding Proteins; D014764:Viral Proteins