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Characterization of the Methionine Sulfoxide Reductase Activities of PILB, a Probable Virulence Factor from Neisseria meningitidis

Abstract : PILB has been described as being involved in the virulence of bacteria of Neisseria genus. The PILB protein is composed of three subdomains. In the present study, the central subdomain (PILB-MsrA), the C terminus subdomain (PILB-MsrB), and the fused subdomain (PILB-MsrA/MsrB) of N. meningitidiswere produced as folded entities. The central subdomain shows a methionine sulfoxide reductase A (MsrA) activity, whereas PILB-MsrB displays a methionine sulfoxide reductase B (MsrB) activity. The catalytic mechanism of PILB-MsrB can be divided into two steps: 1) an attack of the Cys-494 on the sulfur atom of the sulfoxide substrate, leading to formation of a sulfenic acid intermediate and release of 1 mol of methionine/mol of enzyme and 2) a regeneration of Cys-494 via formation of an intradisulfide bond with Cys-439 followed by reduction with thioredoxin. The study also shows that 1) MsrA and MsrB display opposite stereoselectivities toward the sulfoxide function; 2) the active sites of both Msrs, particularly MsrB, are rather adapted for binding protein-bound MetSO more efficiently than free MetSO; 3) the carbon Cα is not a determining factor for efficient binding to both Msrs; and 4) the presence of the sulfoxide function is a prerequisite for binding to Msrs. The fact that the two Msrs exhibit opposite stereoselectivities argues for a structure of the active site of MsrBs different from that of MsrAs. This is further supported by the absence of sequence homology between the two Msrs in particular around the cysteine that is involved in formation of the sulfenic acid derivative. The fact that the catalytic mechanism takes place through formation of a sulfenic acid intermediate for both Msrs supports the idea that sulfenic acid chemistry is a general feature in the reduction of sulfoxides by thiols.
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J. Biol. Chem.-2002-Olry-12016...
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Alexandre Olry, Sandrine Boschi-Muller, Michel Marraud, Sarah Sanglier-Cianferani, Alain van Dorsselear, et al.. Characterization of the Methionine Sulfoxide Reductase Activities of PILB, a Probable Virulence Factor from Neisseria meningitidis. Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2002, 277 (14), pp.12016-12022. ⟨10.1074/jbc.M112350200⟩. ⟨hal-01690847⟩



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