Purification of human active urinary kallikrein: comparative inhibition studies of kininogenase and amidolytic activities.
Large scale purification of human active urinary kallikrein is described. The final preparation was found homogeneous by means of SDS Page electrophoresis, amino acid composition and N-terminal analysis. The apparent molecular weight, determined on SDS Page electrophoresis, was 4.4 X 10(4). Comparative inhibition studies of the kininogenase and the amidase activities pointed out differences in the sensitivity of these two activities. Sodium inhibited amidase activity whereas kininogenase activity required the presence of this cation. In contrast, kininogenase activity was more sensitive to cadmium inhibition than amidase activity. Antibody against purified kallikrein did not completely inhibit amidase activity in crude urine. These discrepancies are consistent with the existence of several amidase activities in urine and also with possibly distinct catalytic sites on the same molecule, accordingly consideration of the methodology used appears very important when comparing results from different studies.
Girolami
J P
JP
INSERM U133 Faculté de Médecine Rangueil, Toulouse, France.
Pecher
C
C
Bascands
J L
JL
Cabos
G
G
Adam
A
A
Suc
J M
JM
D000581:Amidohydrolases / Q000378:metabolism; D000596:Amino Acids / Q000032:analysis; D002104:Cadmium / Q000494:pharmacology; D002852:Chromatography, Ion Exchange; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007610:Kallikreins / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism / Q000652:urine*; D012710:Serum Albumin, Bovine / Q000494:pharmacology
D000596:Amino Acids; D002104:Cadmium; D012710:Serum Albumin, Bovine; D004492:Edetic Acid; D007610:Kallikreins; D000581:Amidohydrolases