pH-sensitive control of arginase by Mn(II) ions at submicromolar concentrations.
The manganese dependence of arginase was reinvestigated with extracts of mouse liver to see whether more physiological properties were displayed than have been reported for the purified enzyme. In a preincubation with Mn(II) ions at 37 degrees C the enzyme underwent a slow and reversible activation. At least 90-95% of the activation achieved was dependent on Mn2+. However, no Mn2+ was required for catalytic activity in the assay. The activation showed little dependence upon pH over the range 6.5-9.5, whereas the catalytic activity increased 12-fold in apparent accord with the titration curve of an ionizable group of pKa 7.9. The Mn2+ dependence of arginase activation obeyed Michaelis-Menten kinetics, with Kd varying from 0.3 microM at pH 6.8 to 0.08 microns at pH 7.7. Free Mn2+ concentrations were established in these assays with a trimethylenediaminetetraacetate-Mn buffer. Vmax increased about three-fold over this range. The calculated arginase activity at 0.05 microM Mn2+ increases about nine-fold over this physiological pH range. An enzyme model is proposed to explain these findings. The activity of arginase at "physiological" [Mn2+] and the pronounced pH dependence conferred upon it are consistent with a recently revised role for the urea cycle in the control of bicarbonate and pH in the body. It appears possible that arginase loses Mn2+ sensitivity during the usual purification.
Kuhn
N J
NJ
School of Biochemistry, University of Birmingham, United Kingdom.
Talbot
J
J
Ward
S
S
D000818:Animals; D001119:Arginase / Q000302:isolation & purification / Q000378:metabolism*; D002712:Chlorides; D004492:Edetic Acid / Q000494:pharmacology; D004789:Enzyme Activation; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008345:Manganese / Q000494:pharmacology*; D017895:Manganese Compounds; D051379:Mice; D008954:Models, Biological
D002712:Chlorides; D017895:Manganese Compounds; D008345:Manganese; D004492:Edetic Acid; D001119:Arginase; C025340:manganese chloride