Arginine residues in the active site of human phenol sulfotransferase (SULT1A1)

Cytosolic sulfotransferases (STs) catalyze the sulfation of hydroxyl containing compounds. Human phenol sulfotransferase (SULT1A1) is the major human ST that catalyzes the sulfation of simple phenols. Because of its broad substrate specificity and lack of endogenous substrates, the biological function of SULT1A1 is believed to be an important detoxification enzyme. In this report, amino acid modification, computer structure modeling, and site-directed mutagenesis were used for studies of Arg residues in the active site of SULT1A1. The Arg-specific modification reagent, 2,3-butanedione, inactivated SULT1A1 in an efficient, time- and concentration-dependent manner, suggesting Arg residues play an important role in the catalytic activity of SULT1A1. According to the computer model, Arg(78), Arg(130), and Arg(257) may be important for SULT1A1 catalytic activity. Site-directed mutagenesis results demonstrated that the positive charge on Arg(78) is not critical for SULT1A1 because R78A is still active. In contrast, a negative charge at this position, R78E, completely inactivated SULT1A1. Arg(78) is in close proximity to the site of sulfuryl group transfer. Arg(257) is located very close to the 3'-phosphate in adenosine 3'-phosphate 5'-phosphosulfate ( PAPS). Site-directed mutagenesis demonstrated that Arg(257) is critical for SULT1A1: both R257A and R257E are inactive. Although Arg(130) is also located very close to the 3'-phosphate of PAPS, R130A and R130E are still active, suggesting that Arg(130) is not a critical residue for the catalytic activity of SULT1A1. Computer modeling suggests that the ionic interaction between the positive charge on Arg(257), and the negative charge on 3'-phosphate is the primary force stabilizing the specific binding of PAPS.