Phosphate is an inhibitor of copper-zinc superoxide dismutase.
The superoxide dismutase (SOD) activity of bovine copper-zinc superoxide dismutase (Cu,Zn-SOD) in 50 mM Hepes [4-(2-hydroxyethyl)-1- piperazineethanesulfonic acid], pH 7.4, was decreased by approximately 50% when the solution was made 10 mM in phosphate, in spite of the fact that the ionic strength of both solutions was adjusted to be equal. A similar experiment was carried out with bovine Cu,Zn-SOD chemically modified at Arg-141 with phenylglyoxal, which consequently had approximately 20% of the activity of the unmodified protein. (This activity was shown not to be due to residual unmodified protein.) Addition of 10 mM phosphate to solutions of the modified protein caused only a small decrease (less than 5%) in the SOD activity. The presence of phosphate also caused the affinity of Cu,Zn-SOD for binding azide or cyanide anions to be reduced; this effect of phosphate was also much less for the arginine-modified protein. We conclude that the inhibitory effect of phosphate on bovine Cu,Zn-SOD is due primarily to the neutralization of the positive charge on the side chain of Arg-141. The effect of increasing ionic strength on the activities of the native and arginine-modified proteins was also investigated. We found that at high concentrations of phosphate (greater than or equal to 10 mM), the SOD activities of native and arginine-modified Cu,Zn-SOD were inhibited comparably when the ionic strength was increased. This effect is presumably due to the lysine residues near the active site.(ABSTRACT TRUNCATED AT 250 WORDS)
D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine; D001386:Azides; D001665:Binding Sites; D002417:Cattle; D003300:Copper / Q000494:pharmacology; D003486:Cyanides; D066298:In Vitro Techniques; D010710:Phosphates / Q000494:pharmacology*; D013482:Superoxide Dismutase / Q000037:antagonists & inhibitors*; D015032:Zinc / Q000494:pharmacology
D001386:Azides; D003486:Cyanides; D010710:Phosphates; D003300:Copper; D001120:Arginine; D013482:Superoxide Dismutase; D015032:Zinc