Reversible methionine sulfoxidation of Mycobacterium tuberculosis small heat shock protein Hsp16.3 and its possible role in scavenging oxidants.
Mycobacterium tuberculosis (TB) small heat shock protein Hsp16.3 was found to be a major membrane protein that is most predominantly expressed under oxidative stress and is localized to the thickened cell envelope. Gene knock-out studies indicate that the Hsp16.3 protein is required for TB to grow in its host macrophage cells. The physiological function of Hsp16.3 has not yet revealed. Our analyses via mass spectrometry, conformation-dependent trypsin digestion, nondenaturing pore gradient electrophoresis, ANS-binding fluorescence measurements, and circular dichroism demonstrate that the three and only the three methionine residues (cysteine and tryptophan residues, which can also be readily oxidized by such oxidant as H(2)O(2), are absent in Hsp16.3) can be readily sulfoxidized with H(2)O(2) treatment in vitro, and the methionine sulfoxide can be effectively reduced back to the methionine form. Interconversion between the methionine and methioninesulfoxide has been confirmed by selective oxidation and reduction. The sulfoxidation leads to a small degree of conformational change, which in turn results in a significant decrease of the chaperone-like activity. Data presented in this report strongly implicate that reversible sulfoxidation/desulfoxidation of methionine residues may occur in Hsp16.3, which serves as a way to scavenger reactive oxygen or nitrogen species abundantly present in macrophage cells, thus protecting the plasma membrane and other components of M. tuberculosis allowing their survival in such bacteriocidal hosts.
Abulimiti
Abuduaini
A
Department of Biological Science and Biotechnology, School of Life Science, Tsinghua University, Beijing, 100084, PR China.
Qiu
Xiaolei
X
Chen
Jing
J
Liu
Yang
Y
Chang
Zengyi
Z
D001426:Bacterial Proteins; D018833:Chaperonins / Q000737:chemistry* / Q000502:physiology*; D006861:Hydrogen Peroxide / Q000494:pharmacology; D013058:Mass Spectrometry; D008715:Methionine / Q000737:chemistry*; D009169:Mycobacterium tuberculosis; D016877:Oxidants / Q000494:pharmacology; D010084:Oxidation-Reduction; D011487:Protein Conformation; D013455:Sulfur / Q000737:chemistry
D001426:Bacterial Proteins; C431133:Hsp16.3 protein, Mycobacterium tuberculosis; D016877:Oxidants; D013455:Sulfur; D008715:Methionine; D006861:Hydrogen Peroxide; D018833:Chaperonins