Purification and characterization of dog urinary kallikrein.

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Abstract
Two forms of N-.alpha.-tosyl-L-arginine methyl ester (TAME) hydrolyzing activities were found in dog urine and one of them was confirmed to be dog urinary kallikrein (DUK, E.C. 3.4.21.8). Mainly by chromatographic techniques, the kallikrein was purified .apprx. 1390-fold from the dialyzed dog urine with the overall yield of 36% of vasodilator activity. The specific activities of the finally purified preparation of DUK were 1250 kallikrein units (KU)/A280 absorbancy at 280 nm of vasodilator activity and 11.3 .mu.mol/min at A280 of TAME esterolytic activity, and the preparation was homogenous on disc gel electrophoresis and ultracentrifugal analysis. The MW of DUK was .apprx. 2.6 to 2.9 .times. 104 by vertical plate polyacrylamide gel electrophoresis, gel filtration on a Sephadex G-100 column and ultracentrifugation, and the isoelectric point of this enzyme was also determined as pI 4.2. Various properties such as pH stability, optimum pH for esterolytic activity and heat stability were similar to those of dog renal kallikrein.