A low pKa cysteine at the active site of mouse methionine sulfoxide reductase A.
Methionine sulfoxide reductase A is an essential enzyme in the antioxidant system which scavenges reactive oxygen species through cyclic oxidation and reduction of methionine and methionine sulfoxide. Recently it has also been shown to catalyze the reverse reaction, oxidizing methionine residues to methionine sulfoxide. A cysteine at the active site of the enzyme is essential for both reductase and oxidase activities. This cysteine has been reported to have a pK(a) of 9.5 in the absence of substrate, decreasing to 5.7 upon binding of substrate. Using three independent methods, we show that the pK(a) of the active site cysteine of mouse methionine sulfoxide reductase is 7.2 even in the absence of substrate. The primary mechanism by which the pK(a) is lowered is hydrogen bonding of the active site Cys-72 to protonated Glu-115. The low pK(a) renders the active site cysteine susceptible to oxidation to sulfenic acid by micromolar concentrations of hydrogen peroxide. This characteristic supports a role for methionine sulfoxide reductase in redox signaling.
Laboratory of Biochemistry, NHLBI, National Institutes of Health, Bethesda, Maryland 20892-8012, USA.
Jung Chae
JC
Lim
James M
JM
Gruschus
Geumsoo
G
Kim
Barbara S
BS
Berlett
Nico
N
Tjandra
Rodney L
RL
Levine
D000818:Animals; D002384:Catalysis; D020134:Catalytic Domain; D003545:Cysteine / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006860:Hydrogen Bonding; D008715:Methionine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051150:Methionine Sulfoxide Reductases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D051379:Mice; D010084:Oxidation-Reduction
D008715:Methionine; D051150:Methionine Sulfoxide Reductases; C027219:methionine sulfoxide reductase; D003545:Cysteine