Broad specificity AhpC-like peroxiredoxin and its thioredoxin reductant in the sparse antioxidant defense system of Treponema pallidum

• Published in: Proceedings of the National Academy of Sciences - PNAS 2010-04, Vol.107 (14), p.6240-6245
• Main Authors: Parsonage, Derek, Desrosiers, Daniel C, Hazlett, Karsten R.O, Sun, Yongcheng, Nelson, Kimberly J, Cox, David L, Radolf, Justin D, Poole, Leslie B
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Antioxidants; Antioxidants - metabolism; Bacteria; Biological Sciences; Enzymes; Eukaryotes; Genes; Genome, Bacterial; Genomics; Molecular Sequence Data; Oxidation-Reduction; Oxidative stress; Peroxides; Peroxiredoxins - chemistry; Peroxiredoxins - metabolism; Rabbits; Ribonucleic acid; RNA; Sequence Alignment; Spirochaetales; Substrate Specificity; Superoxides; Syphilis; Thioredoxin; Thioredoxins - metabolism; Transcription, Genetic; Treponema pallidum; Treponema pallidum - genetics; Treponema pallidum - metabolism
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.0910057107

Little is known about the mechanisms by which Treponema pallidum (Tp), the causative agent of syphilis, copes with oxidative stress as it establishes persistent infection within its obligate human host. The Tp genomic sequence indicates that the bacterium's antioxidant defenses do not include glutathione and are limited to just a few proteins, with only one, TP0509, offering direct defense against peroxides. Although this Tp peroxiredoxin (Prx) closely resembles AhpC-like Prxs, Tp lacks AhpF, the typical reductant for such enzymes. Functionally, TpAhpC resembles largely eukaryotic, nonAhpC typical 2-Cys Prx proteins in using thioredoxin (Trx, TP0919) as an efficient electron donor and exhibiting broad specificity toward hydroperoxide substrates. Unlike many of the eukaryotic Prxs, however, TpAhpC is relatively resistant to inactivation during turnover with hydroperoxide substrates. As is often observed in typical 2-Cys Prxs, TpAhpC undergoes redox-sensitive oligomer formation. Quantitative immunoblotting revealed that TpTrx and TpAhpC are present at very high levels (over 100 and 300 μM, respectively) in treponemes infecting rabbit testes; their redox potentials, at -242 ± 1 and -192 ± 2 mV, respectively, are consistent with the role of TpTrx as the cellular reductant of TpAhpC. Transcriptional analysis of select antioxidant genes confirmed the presence of high mRNA levels for ahpC and trx which diminish greatly when spirochetes replicate under in vitro growth conditions. Thus, T. pallidum has evolved an extraordinarily robust, broad-spectrum AhpC as its sole mechanism for peroxide defense to combat this significant threat to treponemal growth and survival during infection.