The general transition metal (Tro) and Zn2+(Znu) transporters inTreponema pallidum: analysis of metal specificities and expression profiles
| Autor: | Justin D. Radolf, Yong Cheng Sun, David L. Cox, Akbar A. Zaidi, Daniel C. Desrosiers, Christian H. Eggers |
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| Rok vydání: | 2007 |
| Předmět: |
Male
Models Molecular Circular dichroism Protein Conformation ATP-binding cassette transporter Microbiology Metal Protein structure Bacterial Proteins Animals Treponema pallidum Molecular Biology Manganese Treponema biology Gene Expression Profiling Binding protein Isothermal titration calorimetry Gene Expression Regulation Bacterial Periplasmic space biology.organism_classification Molecular biology Zinc Periplasmic Binding Proteins visual_art visual_art.visual_art_medium ATP-Binding Cassette Transporters Rabbits |
| Zdroj: | Molecular Microbiology. 65:137-152 |
| ISSN: | 1365-2958 0950-382X |
| DOI: | 10.1111/j.1365-2958.2007.05771.x |
| Popis: | Acquisition of transition metals is central to the struggle between a bacterial pathogen and its mammalian host. Previous studies demonstrated that Treponema pallidum encodes a cluster-9 (C9) ABC transporter (troABCD) whose solute-binding protein component (TroA) ligands Zn(2+) and Mn(2+) with essentially equal affinities. Bioinformatic analysis revealed that T. pallidum encodes an additional C9 transporter (tp0034-36) orthologous to Zn(2+)-uptake (Znu) systems in other bacteria; the binding protein component, ZnuA, contains a His-rich tract characteristic of C9 Zn(2+)-binding proteins. Metal analysis and metal-reconstitution studies demonstrated that ZnuA is a Zn(2+)-binding protein; parallel studies confirmed that TroA binds Zn(2+), Mn(2+) and Fe. Circular dichroism showed that ZnuA, but not TroA, undergoes conformational changes in the presence of Zn(2+). Using isothermal titration calorimetry (ITC), we demonstrated that TroA binds Zn(2+) and Mn(2+) with affinities approximately 100-fold greater than those previously reported. ITC analysis revealed that ZnuA contains multiple Zn(2+)-binding sites, two of which are high-affinity and presumed to be located within the binding pocket and His-rich loop. Quantitative reverse transcription polymerase chain reaction of tro and znu transcripts combined with immunoblot analysis of TroA and ZnuA confirmed that both transporters are simultaneously expressed in T. pallidum and that TroA is expressed at much greater levels than ZnuA. Collectively, our findings indicate that T. pallidum procures transition metals via the concerted utilization of its general metal (Tro) and Zn(2+) (Znu) transporters. Sequestration of periplasmic Zn(2+) by ZnuA may free up TroA binding capacity for the importation of Fe and Mn(2+). |
| Databáze: | OpenAIRE |
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