title: Three different putative phosphate transport receptors are encoded by the Mycobacterium tuberculosis genome and are present at the surface of Mycobacterium bovis BCG.
abstract: A gene encoding a protein homologous to the periplasmic ABC phosphate binding receptor PstS from Escherichia coli was cloned and sequenced from a lambda gt11 library of Mycobacterium tuberculosis by screening with monoclonal antibody 2A1-2. Its degree of similarity to the E. coli PstS is comparable to those of the previously described M. tuberculosis phosphate binding protein pab (Ag78, Ag5, or 38-kDa protein) and another M. tuberculosis protein which we identified recently. We suggest that the three M. tuberculosis proteins share a similar function and could be named PstS-1, PstS-2, and PstS-3, respectively. Molecular modeling of their three-dimensional structures using the structure of the E. coli PstS as a template and their inducibility by phosphate starvation support this view. Recombinant PstS-2 and PstS-3 were produced and purified by affinity chromatography. With PstS-1, these proteins were used to demonstrate the specificity of three groups of monoclonal antibodies. Using these antibodies in flow cytometry and immunoblotting analyses, we demonstrate that the three genes are expressed and their protein products are present and accessible at the mycobacterial surface as well as in its culture filtrate. Together with the M. tuberculosis genes encoding homologs of the PstA, PstB, and PstC components we cloned before, the present data suggest that at least one, and possibly several, related and functional ABC phosphate transporters exist in mycobacteria. It is hypothesized that the mycobacterial gene duplications presented here may be a subtle adaptation of intracellular pathogens to phosphate starvation in their alternating growth environments.
authors.affiliation: Department of Virology, Institut Pasteur, Brussels, Belgium.
authors.forename: P
authors.initials: P
authors.lastname: Lefèvre
authors.forename: M
authors.initials: M
authors.lastname: Braibant
authors.forename: L
authors.initials: L
authors.lastname: de Wit
authors.forename: M
authors.initials: M
authors.lastname: Kalai
authors.forename: D
authors.initials: D
authors.lastname: Röeper
authors.forename: J
authors.initials: J
authors.lastname: Grötzinger
authors.forename: J P
authors.initials: JP
authors.lastname: Delville
authors.forename: P
authors.initials: P
authors.lastname: Peirs
authors.forename: J
authors.initials: J
authors.lastname: Ooms
authors.forename: K
authors.initials: K
authors.lastname: Huygen
authors.forename: J
authors.initials: J
authors.lastname: Content
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002352:Carrier Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D017931:DNA Primers; D029968:Escherichia coli Proteins; D016680:Genome, Bacterial; D008565:Membrane Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000145:classification / Q000235:genetics; D009163:Mycobacterium bovis / Q000235:genetics* / Q000378:metabolism*; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000378:metabolism*; D033902:Periplasmic Binding Proteins; D028044:Phosphate-Binding Proteins; D010710:Phosphates / Q000378:metabolism; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
chemical_list: D002352:Carrier Proteins; D017931:DNA Primers; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D033902:Periplasmic Binding Proteins; D028044:Phosphate-Binding Proteins; D010710:Phosphates; C090133:PstS protein, E coli; D011994:Recombinant Proteins