STUDIES ON RAT RENAL CORTICAL CELL KALLIKREIN .2. IDENTIFICATION OF KALLIKREIN AS AN ECTO-ENZYME

被引:41
作者
CHAO, J [1 ]
MARGOLIUS, HS [1 ]
机构
[1] MED UNIV S CAROLINA,DEPT MED,CHARLESTON,SC 29403
关键词
(Rat renal cortex); Cell surface; Ectoenzyme; Kallikrein;
D O I
10.1016/0005-2744(79)90153-0
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Suspensions of viable renal cortical cells hydrolyzed a synthetic ester substrate (α-N-tosyl-spl-argnine, methyl ester, Tos-Arg-OMe) and generated kinins from a kininogen substrate. This kallikrein-like esterase activity increased linearly with cell number, or time of exposure to substrate. No radiolabelled substrate or product was found within the cells. Most of the activity appeared to be on cell surfaces as supernatant media had less than 20% of the Tos-Arg-OMe esterase activity of the cell suspensions. Cell surface Tos-Arg-OMe esterase activity was inhibited by aprotinin, benzamidine, pentamidine, and a trisamidine derivative (α,α′,α″-tris(3-amidinophenoxy)mesitylene). Preincubation of cells with phospholipase A2 increased renal cell surface esterase activity up to 76% while only slightly increasing supernatant activity. In contrast, preincubation with deoxycholate caused clearing of suspensions and a marked increase in supernatant esterase activity. Renal cell kininogenase (EC 3.4.21.8) activity was inhibited by preincubation with aprotinin, the tris-amidine derivative, or anti-rat urinary kallikrein antibody. Kallikrein elaborated by renal cells formed a single precipitin line with an antibody to rat urinary kallikrein but the two enzymes were not immunologically identical. We conclude that kallikrein's active sites are facing the external environment of renal cortical cells in suspension with access to substrates, inhibitors, and antibody. © 1979.
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页码:330 / 340
页数:11
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