Structure analysis of the extracellular domain reveals disulfide bond forming-protein properties of <italic>Mycobacterium tuberculosis</italic> Rv2969c

doi:10.1007/s13238-013-3033-x

Prot Cell

2013, Vol. 4

Issue (8) : 628-640 https://doi.org/10.1007/s13238-013-3033-x PMID: 23828196

RESEARCH ARTICLE

Structure analysis of the extracellular domain reveals disulfide bond forming-protein properties of Mycobacterium tuberculosis Rv2969c

Lu Wang^1,2, Jun Li^1,2, Xiangxi Wang^1,2, Wu Liu¹, Xuejun C. Zhang¹, Xuemei Li¹, Zihe Rao^1,3(

)

1. National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; 2. University of Chinese Academy of Sciences, Beijing, China; 3. Tianjin Key Laboratory of Protein Science, College of Life Sciences, Nankai University, Tianjin 300071, China

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Abstract

Disulfide bond-forming (Dsb) protein is a bacterial periplasmic protein that is essential for the correct folding and disulfide bond formation of secreted or cell wallassociated proteins. DsbA introduces disulfide bonds into folding proteins, and is re-oxidized through interaction with its redox partner DsbB. Mycobacterium tuberculosis, a Gram-positive bacterium, expresses a DsbA-like protein ( Rv2969c), an extracellular protein that has its Nterminus anchored in the cell membrane. Since Rv2969c is an essential gene, crucial for disulfide bond formation, research of DsbA may provide a target of a new class of anti-bacterial drugs for treatment of M.tuberculosis infection. In the present work, the crystal structures of theextracellular region of Rv2969c (Mtb DsbA) were determined in both its reduced and oxidized states. The overall structure of Mtb DsbA can be divided into two domains: a classical thioredoxin-like domain with a typical CXXC active site, and an α-helical domain. It largely resembles its Escherichiacoli homologue EcDsbA, however, it possesses a truncated binding groove; in addition, its active site is surrounded by an acidic, rather than hydrophobic surface. In our oxidoreductase activity assay, Mtb DsbA exhibited a different substrate specificity when compared to EcDsbA. Moreover, structural analysis revealed a second disulfide bond in Mtb DsbA, which is rare in the previously reported DsbA structures, and is assumed to contribute to the overall stability of Mtb DsbA. To investigate the disulphide formation pathway in M.tuberculosis, we modeled Mtb Vitamin K epoxide reductase (Mtb VKOR), a binding partner of Mtb DsbA, to Mtb DsbA.

Keywords Mycobacterium tuberculosis disulfide bond forming protein X-ray crystallography

Corresponding Author(s): Rao Zihe,Email:raozh@xtal.tsinghua.edu.cn

Issue Date: 01 August 2013

Cite this article:

Lu Wang,Jun Li,Xiangxi Wang, et al. Structure analysis of the extracellular domain reveals disulfide bond forming-protein properties of Mycobacterium tuberculosis Rv2969c[J]. Prot Cell, 2013, 4(8): 628-640.

URL:

https://academic.hep.com.cn/pac/EN/10.1007/s13238-013-3033-x
https://academic.hep.com.cn/pac/EN/Y2013/V4/I8/628

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