title: The PE-PGRS glycine-rich proteins of Mycobacterium tuberculosis: a new family of fibronectin-binding proteins?
abstract: A clone was isolated by screening of a cosmid library of Mycobacterium tuberculosis with an oligonucleotide designed from the N-terminal sequence of a previously reported proline-rich protein. Characterization of the 4481 bp insert showed the presence of polymorphic CG-repetitive sequences (PGRSs) with an ORF of 2.7 kb, encoding a 81.3 kDa protein (PE-PGRS81). Southern blot analysis and BLAST-p searches revealed several homologous sequences in the genome of M. tuberculosis. The deduced amino acid sequence was highly similar to a stretch of about 98 residues in the N-terminus present in several members of the PE-PGRS family available in the GenBank database, including 100% identity with the partial amino acid sequence of the potential protein encoded by orf3' as well as with the Rv0278c sequence. A neighbour-joining analysis of the 99 PE-PGRS sequences available in the database indicated that PE-PGRS81 is included in a group where its closest relatives are the sequences orf3', Rv0278c, Rv0279c, Rv1759c, Rv3652 and Rv0747. Probing with the complete coding regions of PE-PGRS81 and Rv1759c in Southern blot assays, on samples of genomic DNA from M. tuberculosis H37Rv, Mycobacterium bovis BCG and M. tuberculosis clinical isolates, showed a complex hybridization pattern for all strains. This shows the existence of intrastrain PGRS variability as reported for other PGRS members. In contrast, probing with the short conserved N-terminal region of Rv1759c reduced the hybridization to a single band. This marker allowed identification of M. tuberculosis clinical strains that lack Rv1759c. A recombinant C-terminal fragment of Rv1759c showed fibronectin-binding properties and was recognized by sera from patients infected with M. tuberculosis, suggesting that at least this member of the PE-PGRS is expressed in tuberculosis infection.
authors.affiliation: Departamento de Inmunologı́ a, Instituto de Investigaciones Biomé dicas, UNAM, Apartado Postal 70228, 04510 México DF, Mexico1.
authors.forename: Clara
authors.initials: C
authors.lastname: Espitia
authors.affiliation: Departamento de Inmunologı́ a, Instituto de Investigaciones Biomé dicas, UNAM, Apartado Postal 70228, 04510 México DF, Mexico1.
authors.forename: Juan Pedro
authors.initials: JP
authors.lastname: Laclette
authors.affiliation: Departamento de Inmunologı́ a, Instituto de Investigaciones Biomé dicas, UNAM, Apartado Postal 70228, 04510 México DF, Mexico1.
authors.forename: Mariana
authors.initials: M
authors.lastname: Mondragón-Palomino
authors.affiliation: Departamento de Inmunologı́ a, Instituto de Investigaciones Biomé dicas, UNAM, Apartado Postal 70228, 04510 México DF, Mexico1.
authors.forename: Angelica
authors.initials: A
authors.lastname: Amador
authors.affiliation: Departamento de Inmunologı́ a, Instituto de Investigaciones Biomé dicas, UNAM, Apartado Postal 70228, 04510 México DF, Mexico1.
authors.forename: Jaime
authors.initials: J
authors.lastname: Campuzano
authors.affiliation: GBF, German National Research Center for Biotechnology, 38124 Braunschweig, Germany2.
authors.forename: Anke
authors.initials: A
authors.lastname: Martens
authors.affiliation: GBF, German National Research Center for Biotechnology, 38124 Braunschweig, Germany2.
authors.forename: Mahavir
authors.initials: M
authors.lastname: Singh
authors.affiliation: Hospital General de México, SSA , Mexico3.
authors.forename: Raul
authors.initials: R
authors.lastname: Cicero
authors.affiliation: MRC Tuberculosis and Related Infections Unit, Clinical Sciences Centre, Hammersmith Hospital, Du Cane Road, London, UK 4.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: MRC Tuberculosis and Related Infections Unit, Clinical Sciences Centre, Hammersmith Hospital, Du Cane Road, London, UK 4.
authors.forename: Carlos
authors.initials: C
authors.lastname: Moreno
mesh_terms: D018829:Adhesins, Bacterial; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D015139:Blotting, Southern; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004269:DNA, Bacterial / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005353:Fibronectins / Q000378:metabolism*; D005998:Glycine / Q000737:chemistry; D006801:Humans; D015151:Immunoblotting; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000378:metabolism*; D010802:Phylogeny; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length; D011994:Recombinant Proteins / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014376:Tuberculosis / Q000382:microbiology; D014397:Tuberculosis, Pulmonary / Q000382:microbiology
chemical_list: D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; D002352:Carrier Proteins; D004269:DNA, Bacterial; D005353:Fibronectins; D011994:Recombinant Proteins; C078635:fibronectin-binding proteins, bacterial; D005998:Glycine