title: Molecular function of WhiB4/Rv3681c of Mycobacterium tuberculosis H37Rv: a [4Fe-4S] cluster co-ordinating protein disulphide reductase.
abstract: The genome sequence of Mycobacterium tuberculosis H37Rv revealed the presence of seven whiB-like open reading frames. In spite of several genetic studies on whiB genes, the biochemical properties of WhiB proteins are poorly understood. All WhiB-like proteins have four conserved cysteine residues, out of which two are present in a CXXC motif. We report for the first time the detailed biochemical and biophysical properties of M. tuberculosis WhiB4/Rv3681c and demonstrate the functional relevance of four conserved cysteines and the CXXC motif. UV-visible absorption spectra of freshly purified mWhiB4 showed the presence of a [2Fe-2S] cluster, whereas the electron paramagnetic resonance (EPR) spectra of reconstituted protein showed the presence of a [4Fe-4S] cluster. The iron-sulphur cluster was redox sensitive but stably co-ordinated to the protein even in the presence of high concentration of chaotropic agents. Despite primary sequence divergence from thioredoxin family proteins, the apo mWhiB4 has properties similar to thioredoxins and functions as a protein disulphide reductase, whereas holo mWhiB4 is enzymatically inactive. Apart from the cysteine thiol of CXXC motif the distantly placed thiol pair also contributes equally to the enzymatic activity of mWhiB4. A functional model of mWhiB4 in redox signaling during oxidative stress in M. tuberculosis has been presented.
authors.affiliation: Institute of Microbial Technology, Sector-39A, Chandigarh, 160 036, India.
authors.forename: Md Suhail
authors.initials: MS
authors.lastname: Alam
authors.forename: Saurabh K
authors.initials: SK
authors.lastname: Garg
authors.forename: Pushpa
authors.initials: P
authors.lastname: Agrawal
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D004578:Electron Spin Resonance Spectroscopy; D007501:Iron / Q000032:analysis; D008027:Light; D008954:Models, Biological; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000201:enzymology*; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010088:Oxidoreductases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D016415:Sequence Alignment; D013057:Spectrum Analysis; D013455:Sulfur / Q000032:analysis; D013880:Thioredoxin-Disulfide Reductase / Q000378:metabolism; D014466:Ultraviolet Rays
chemical_list: D013455:Sulfur; D007501:Iron; D010088:Oxidoreductases; D013880:Thioredoxin-Disulfide Reductase