Structural and functional characterization of the Pseudomonas hydroperoxide resistance protein Ohr

被引:79
作者
Lesniak, J
Barton, WA
Nikolov, DB
机构
[1] Cornell Univ, Joan & Sanford I Weill Grad Sch Med Sci, New York, NY 10021 USA
[2] Mem Sloan Kettering Canc Ctr, Cellular Biochem & Biophys Program, New York, NY 10021 USA
关键词
bacterial resistance; organic hydroperoxides; oxidative stress; peroxiredoxin; Pseudomonas aeruginosa;
D O I
10.1093/emboj/cdf670
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Bacteria have developed complex strategies to detoxify and repair damage caused by reactive oxygen species. These compounds, produced during bacterial aerobic respiration as well as by the host immune system cells as a defense mechanism against the pathogenic microorganisms, have the ability to damage nucleic acids, proteins and phospholipid membranes. Here we describe the crystal structure of Pseudomonas aeruginosa Ohr, a member of a recently discovered family of organic hydroperoxide resistance proteins. Ohr is a tightly folded homodimer, with a novel alpha/beta fold, and contains two active sites located at the monomer interface on opposite sides of the molecule. Using in vitro assays, we demonstrate that Ohr functions directly as a hydroperoxide reductase, converting both inorganic and organic hydroperoxides to less toxic metabolites. Site-directed mutagenesis confirms that the two conserved cysteines in each active site are essential for catalytic activity. We propose that the Ohr catalytic mechanism is similar to that of the structurally unrelated peroxiredoxins, directly utilizing highly reactive cysteine thiol groups to elicit hydroperoxide reduction.
引用
收藏
页码:6649 / 6659
页数:11
相关论文
共 55 条
[31]   THIOREDOXIN - A FOLD FOR ALL REASONS [J].
MARTIN, JL .
STRUCTURE, 1995, 3 (03) :245-250
[32]   Raster3D: Photorealistic molecular graphics [J].
Merritt, EA ;
Bacon, DJ .
MACROMOLECULAR CRYSTALLOGRAPHY, PT B, 1997, 277 :505-524
[33]   Identification and characterization of a new organic hydroperoxide resistance (ohr) gene with a novel pattern of oxidative stress regulation from Xanthomonas campestris pv. phaseoli [J].
Mongkolsuk, S ;
Praituan, W ;
Loprasert, S ;
Fuangthong, M ;
Chamnongpol, S .
JOURNAL OF BACTERIOLOGY, 1998, 180 (10) :2636-2643
[34]   Purification and characterization of recombinant catalase-peroxidase, which confers isoniazid sensitivity in Mycobacterium tuberculosis [J].
Nagy, JM ;
Cass, AEG ;
Brown, KA .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (50) :31265-31271
[35]   THE EFFECT OF PHENOLIC GLYCOLIPID-1 FROM MYCOBACTERIUM-LEPRAE ON THE ANTIMICROBIAL ACTIVITY OF HUMAN MACROPHAGES [J].
NEILL, MA ;
KLEBANOFF, SJ .
JOURNAL OF EXPERIMENTAL MEDICINE, 1988, 167 (01) :30-42
[36]   PROTEIN FOLDING AND ASSOCIATION - INSIGHTS FROM THE INTERFACIAL AND THERMODYNAMIC PROPERTIES OF HYDROCARBONS [J].
NICHOLLS, A ;
SHARP, KA ;
HONIG, B .
PROTEINS-STRUCTURE FUNCTION AND BIOINFORMATICS, 1991, 11 (04) :281-296
[37]  
NIKI E, 1992, ORGANIC PEROXIDES, P765
[38]   Genetic and physiological characterization of ohr, encoding a protein involved in organic hydroperoxide resistance in Pseudomonas aeruginosa [J].
Ochsner, UA ;
Hassett, DJ ;
Vasil, ML .
JOURNAL OF BACTERIOLOGY, 2001, 183 (02) :773-778
[39]   Processing of X-ray diffraction data collected in oscillation mode [J].
Otwinowski, Z ;
Minor, W .
MACROMOLECULAR CRYSTALLOGRAPHY, PT A, 1997, 276 :307-326
[40]   Automated protein model building combined with iterative structure refinement [J].
Perrakis, A ;
Morris, R ;
Lamzin, VS .
NATURE STRUCTURAL BIOLOGY, 1999, 6 (05) :458-463