**7. Acknowledgments**

This work was supported by grants from the Howard Hughes Medical Institute (HHMI), NIH and CSIC, Universidad de la República, Uruguay, to RR. MT was partially supported by PEDECIBA Biología, Uruguay. MH was partially supported by a fellowship from ANII, Uruguay. RR is an International Research Scholar of the HHMI. We thank Lucía Piacenza (Facultad de Medicina, Universidad de la República, Uruguay) for her kindly help with the artworks.

#### **8. References**

306 Understanding Tuberculosis – Deciphering the Secret Life of the Bacilli

Fig. 2. **Cellular localization and reducing substrates of peroxidases from** *M. tuberculosis.*  The five Prxs and the heme peroxidase KatG have distinct, although overlapping cellular distributions. *Mt*KatG (orange) has been found in the cytosol, membrane and extracellular space. *Mt*AhpC (blue) is a cytosolic enzyme also that was also found associated to the bacterial membrane. *Mt*TPx (green) was detected in culture media repeatedly. It has also been found in membrane fractions and in the cytosol. *Mt*AhpE (violet), and the putative BcpB and Bcp (yellow) were detected in cell membrane fractions, and the latter also in the cytosol. Reducing systems for *Mt*AhpC and *Mt*Tpx (in grey) are shown without considering their cellular localization. *Mt*AhpE and *Mt*Bcps reducing substrates are still unknown.

*M. tuberculosis* is an extremely successful pathogen, despite of being exposed to cytotoxic peroxides formed inside the phagosome of activated macrophages, its primary host cells. The bacterium expresses a heme-dependent peroxidase, KatG, and various thioldependent peroxidases of the Prx type. From the data reviewed herein, it becomes clear that Prxs from *M. tuberculosis* differ in cellular location, and have diverse oxidizing and reducing substrate specificities, that may explain in part the presence of different subfamilies of Prxs in the bacterium. Available data indicate that at least two of them (*Mt*AhpC and *Mt*TPx) play a role in pathogenesis. The third one, *Mt*AhpE, has an outstanding reactivity with fatty-acid derived hydroperoxides, but since natural reducing substrate(s) has not been identified so far, its peroxidase catalytic activity *in vivo* remains to be confirmed. Similarly, further investigation is required to characterize the two

This work was supported by grants from the Howard Hughes Medical Institute (HHMI), NIH and CSIC, Universidad de la República, Uruguay, to RR. MT was partially supported by PEDECIBA Biología, Uruguay. MH was partially supported by a fellowship from ANII, Uruguay. RR is an International Research Scholar of the HHMI. We thank Lucía Piacenza

**6. Conclusions** 

putative Bcp proteins from *M. tuberculosis.*

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**0**

**17**

*Japan*

**Characteristic Conformational Behaviors**

Mycobacterial mycolic acid (MA) are long chain 2-alkyl branched, 3-hydroxy fatty acid with two intra-chain groups in the so-called meromycolate chain. On the basis of the nature of the functional groups in the meromycolate chains, MAs are categorized into three major groups: *α*-MA with no oxygen-containing intra-chain groups, methoxy-MA (MeO-MA) in which the distal group has a methoxy gorup and Keto-MA in which the distal group has a carbonyl group (Fig. 1) (Watanabe et al., 2001; 2002). MAs are characteristic components of mycobacterial cell envelopes, where a major proportion are covalently bonded to the underlying cell wall arabinogalactan (Goren & Brennan, 1979; McNeil et al., 1991; Minnikin,

In the structural models of the mycobacterial cell envelope proposed previously (Minnikin, 1982; Rastogi, 1991), MAs covalently linked to penta-arabinosyl residues of cell wall arabinogalactan are arranged perpendicular to the cell wall, forming a highly structured monolayer. Recent computer simulation work supported such arrangement of MAs as proposed in the model (Hong & Hopfinger, 2004). This outer leaflet of mycobacterial cell envelope is considered to provide the cells with a special permeability barrier responsible for various physiological and pathogenic features of mycobacterial cells (Daffé et al., 1999). There are various other lipids in the mycobacterial cell envelope and they may also take part in the permeability function of the cell envelope as suggested (Minnikin, 1982; Puech et al., 2001; Rastogi, 1991). Recently, a *Mycobacterium tuberculosis* (*M. tb*) mutant whose MA comprises only *α*-MA (Dubnau et al., 2000), a recombinant mutant having over-produced MeO-MA with no Keto-MA (Yuan et al., 1998) and a mutant having 40 % less cell wall mycolate (Daffé et al., 1999) have been described. These results show that *M. tb* can be viable with highly modified mycolic acid composition and that its pathogenicity may be related to the types of MAs. Those papers also suggest that MAs on the cell envelope have determining effect on the permeability barrier function of the cell wall outer hydrophobic layer barrier and different

MAs may contribute to the cell wall permeability barrier functions in different ways.

In very early studies (Staellberg-Stenhagen & Stenhagen, 1945), the multi-component nature of mycolic acids was not yet known, but it was shown that the total MA formed a stable monolayer on the water surface. It was concluded that MA had extended linear structures, a feature later confirmed by structural analysis (Minnikin, 1982; Minnikin et al., 2002; Rastogi,

**1. Introduction**

1982).

**of Representative Mycolic Acids in**

*Graduate School of Science and Engineering, Saitama University*

**the Interfacial Monolayer**

Masumi Villeneuve

of the catalytic site cysteine to cysteine-sulfinic acid. *J Biol Chem* 277(41): 38029- 38036.

