Enzymatic reduction of arsenic compounds in mammalian systems: the rate-limiting enzyme of rabbit liver arsenic biotransformation is MMA(V) reductase

• Published in: Chemical research in toxicology Vol. 12; no. 12; pp. 1278 - 1283
• Main Authors: Zakharyan, R A, Aposhian, H V
• Format: Journal Article
• Language: English
• Published: United States 01-12-1999
• Subjects: Animals; Arsenicals - pharmacokinetics; Biotransformation; Humans; Hydrogen-Ion Concentration; Liver - enzymology; Male; Metabolic Clearance Rate; Methyltransferases - isolation & purification; Methyltransferases - metabolism; Oxidation-Reduction; Rabbits; Substrate Specificity
• ISSN: 0893-228X
• DOI: 10.1021/tx9901231

A unique enzyme, MMA(V) reductase, has been partially purified from rabbit liver by using DEAE-cellulose, carboxymethylcellulose, and red dye ligand chromatography. The enzyme is unique since it is the rate-limiting enzyme in the biotransformation of inorganic arsenite in rabbit liver. The K(m) and V(max) values were 2.16 x 10(-)(3) M and 10.3 micromol h(-)(1) (mg of protein)(-)(1). When DMA(V) or arsenate was tested as a substrate, the K(m) was 20.9 x 10(-)(3) or 109 x 10(-)(3) M, respectively. The enzyme has an absolute requirement for GSH. Other thiols such as DTT or L-cysteine were inactive alone. At a pH below the physiological pH, GSH carried out this reduction, but this GSH reduction in the absence of the enzyme had little if any value at pH 7.4. When the K(m) values of rabbit liver arsenite methyltransferase (5.5 x 10(-)(6) M) and MMA(III) methyltransferase (9.2 x 10(-)(6)) were compared to that of MMA(V) reductase (2.16 x 10(-)(3) M), it can be concluded that MMA(V) reductase was the rate-limiting enzyme of inorganic arsenite biotransformation. MMA(V) reductase was also present in surgically removed human liver.