title: Characterising Mycobacterium tuberculosis Rv1510c protein and determining its sequences that specifically bind to two target cell lines.
abstract: The process of Mycobacterium tuberculosis infection of the macrophage implies a very little-known initial recognition and adherence step, important for mycobacterial survival; many proteins even remain like hypothetical. The Rv1510c gene, encoding a putatively conserved membrane protein, was investigated by analysing the M. tuberculosis genome sequence data reported by Cole et al. and a previous report that used PCR assays to show that the Rv1510 gene was only present in M. tuberculosis. This article confirmed all the above and identified the transcribed gene in M. tuberculosis, Mycobacterium africanum, and in M. tuberculosis clinical isolates. Antibodies raised against peptides from this protein recognised a 44 kDa band, corresponding to Rv1510c theoretical mass (44,294 Da). Assays involving synthetic peptides covering the whole protein binding to U937 and A549 cell lines led to recognising five high activity binding peptides in the Rv1510 protein: 11094, 11095, 11105, 11108, and 11111. Their affinity constants and Hill coefficients were determined by using U937 cells. Cross-linking assays performed with some of these HABPs showed that they specifically bound to a U937 cell line 51 kDa protein, but not to Hep G2 or red blood cell proteins, showing this interaction's specificity.
authors.affiliation: Fundación Instituto de Inmunología de Colombia, Universidad Nacional de Colombia, Carrera 50, No 26-00, Bogotá, Colombia. rivebra@hotmail.com
authors.forename: Ricardo
authors.initials: R
authors.lastname: Vera-Bravo
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Torres
authors.forename: John J
authors.initials: JJ
authors.lastname: Valbuena
authors.forename: Marisol
authors.initials: M
authors.lastname: Ocampo
authors.forename: Luis E
authors.initials: LE
authors.lastname: Rodríguez
authors.forename: Alvaro
authors.initials: A
authors.lastname: Puentes
authors.forename: Javier E
authors.initials: JE
authors.lastname: García
authors.forename: Hernando
authors.initials: H
authors.lastname: Curtidor
authors.forename: Jimena
authors.initials: J
authors.lastname: Cortés
authors.forename: Magnolia
authors.initials: M
authors.lastname: Vanegas
authors.forename: Zuly J
authors.initials: ZJ
authors.lastname: Rivera
authors.forename: Andrea
authors.initials: A
authors.lastname: Díaz
authors.forename: Martha N
authors.initials: MN
authors.lastname: Calderon
authors.forename: Manuel A
authors.initials: MA
authors.lastname: Patarroyo
authors.forename: Manuel E
authors.initials: ME
authors.lastname: Patarroyo
mesh_terms: D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D002942:Circular Dichroism; D003432:Cross-Linking Reagents; D004269:DNA, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D006801:Humans; D007700:Kinetics; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020298:U937 Cells
chemical_list: D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D003432:Cross-Linking Reagents; D004269:DNA, Bacterial; D008565:Membrane Proteins; D010446:Peptide Fragments