Structural and biochemical characterization of the essential DsbA-like disulfide bond forming protein from Mycobacterium tuberculosis

BMC Structural Biology

Celia W Goulding²  David J Guzman²  Christine A Harmston¹  Nicholas Chim¹ 

[1] Departments of Molecular Biology and Biochemistry, UCI, Irvine, CA 92697, USA;Pharmaceutical Sciences, UCI, Irvine, CA 92697, USA

关键词: Oxidoreductase;    Vitamin K epoxide reductase;    DsbA;    X-ray crystallography;    Disulfide bond;    Mycobacterium tuberculosis;   

Others  :  793773
DOI  :  10.1186/1472-6807-13-23

 received in 2013-07-09, accepted in 2013-10-11,  发布年份 2013

【 摘 要 】

Background

Bacterial Disulfide bond forming (Dsb) proteins facilitate proper folding and disulfide bond formation of periplasmic and secreted proteins. Previously, we have shown that Mycobacterium tuberculosis Mt-DsbE and Mt-DsbF aid in vitro oxidative folding of proteins. The M. tuberculosis proteome contains another predicted membrane-tethered Dsb protein, Mt-DsbA, which is encoded by an essential gene.

Results

Herein, we present structural and biochemical analyses of Mt-DsbA. The X-ray crystal structure of Mt-DsbA reveals a two-domain structure, comprising a canonical thioredoxin domain with the conserved CXXC active site cysteines in their reduced form, and an inserted α-helical domain containing a structural disulfide bond. The overall fold of Mt-DsbA resembles that of other DsbA-like proteins and not Mt-DsbE or Mt-DsbF. Biochemical characterization demonstrates that, unlike Mt-DsbE and Mt-DsbF, Mt-DsbA is unable to oxidatively fold reduced, denatured hirudin. Moreover, on the substrates tested in this study, Mt-DsbA has disulfide bond isomerase activity contrary to Mt-DsbE and Mt-DsbF.

Conclusion

These results suggest that Mt-DsbA acts upon a distinct subset of substrates as compared to Mt-DsbE and Mt-DsbF. One could speculate that Mt-DsbE and Mt-DsbF are functionally redundant whereas Mt-DsbA is not, offering an explanation for the essentiality of Mt-DsbA in M. tuberculosis.

【 授权许可】

   
2013 Chim et al.; licensee BioMed Central Ltd.

【 图 表 】

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