Isolation and characterization of rat plasma glandular kallikrein.

A method has been developed to purify glandular kallikrein present in rat plasma by using Sepharose-Aprotinin affinity chromatography and elution of the enzyme with p-aminobenzamidine. The isolated enzyme liberated kinins from kininogen II of low molecular weight (sp. act. 14 ng kinins/min X mg) and p-nitroaniline (pNA) from the substrate S-2266 (sp. act. 1.23 nmoles pNA/min X mg); it was inhibited by aprotinin, benzamidine and rat urinary antikallikrein antibody but not by ovomucoid. In polyacrylamide gel electrophoresis, the enzymatic activities of the preparation were associated with two light protein bands of molecular weights equal to that of urinary kallikrein (35,000 daltons). Using this method, the recovery of [125I]kallikrein added to the plasma was 82-88%. The concentration of the enzyme in normal rat plasma was equivalent to 6.1 +/- 2.1 (S.D.) ng kallikrein/ml. The mean value found in nephrectomized rats was 20.0 +/- 6.3 (S.D.) ng kallikrein/ml. This increment was highly significant (P less than 0.001). Our results confirm the presence of glandular kallikrein in plasma which had been detected by other methods; they also demonstrate that the material purified from plasma is enzymatically active, suggesting that kallikrein may play a biological role in the control of blood circulation.