Isolation and Characterization of Monkey Kidney Prostaglandin-9-Keto Reductase

Abstract

The enzyme, prostaglandin-9-keto reductase, has been partially purified from monkey kidney tissue. DEAE cellulose column chromatography and hydroxylapatite column chromatography were the two procedures used to purify the enzyme. Eleven-fold purification and 47% recovery of the enzyme were achieved. A simple assay was developed using spectrophotometric measurements of the disappearance of NADPH. Substrate specificity of the enzyme was found to be very broad. Changes made in the upper chain and the lower chain of the prostaglandin, effect little change in the activity of the enzyme. Changes made within the ring of the prostaglandin result in loss of activity of the enzyme. Kidney vaso-active agents, such as bradykinin, angiotensin II, and ethacrynic acid, produced no effect on the activity of the enzyme.