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<i>Paracoccus denitrificans</i> ArsH is encoded by two identical genes located in two distinct putative arsenic resistance (<i>ars</i>) operons. <i>Escherichia coli</i>-produced recombinant N-His₆-ArsH was characterized both structurally and kinetically. The X-ray structure of ArsH revealed a flavodoxin-like domain and motifs for the binding of flavin mononucleotide (FMN) and reduced nicotinamide adenine dinucleotide phosphate (NADPH). The protein catalyzed FMN reduction by NADPH via ternary complex mechanism. At a fixed saturating FMN concentration, it acted as an NADPH-dependent organoarsenic reductase displaying ping-pong kinetics. A 1:1 enzymatic reaction of phenylarsonic acid with the reduced form of FMN (FMNH₂) and formation of phenylarsonous acid were observed. Growth experiments with <i>P. denitrificans</i> and <i>E. coli</i> revealed increased toxicity of phenylarsonic acid to cells expressing <i>arsH</i>, which may be related to in vivo conversion of pentavalent As to more toxic trivalent form. ArsH expression was upregulated not only by arsenite, but also by redox-active agents paraquat, tert-butyl hydroperoxide and diamide. A crucial role is played by the homodimeric transcriptional repressor ArsR, which was shown in in vitro experiments to monomerize and release from the DNA-target site. Collectively, our results establish ArsH as responsible for enhancement of organo-As(V) toxicity and demonstrate redox control of <i>ars</i> operon.