Guder W G, Hallbach J, Fink E, Kaissling B, Wirthensohn G
Biol Chem Hoppe Seyler. 1987 Jun;368(6):637-45. doi: 10.1515/bchm3.1987.368.1.637.
Kininogenase activity of kallikrein was measured in microdissected mouse nephron segments using kininogen from dog plasma and a radioimmunoassay for bradykinin. When single nephron segments were examined, results showed a large scatter. This was found to be due to heterogeneity of distal convoluted tubules (DCT) from different nephrons, since replicate measurements in pools of DCT structures did not show this degree of variation. Nearly 20% of activity was accessible to extracellular substrate when freshly dissected segments were incubated in isoosmotic media. Freezing and thawing which markedly releases activity of intracellular enzymes, did not significantly elevate kininogenase activity. On the other hand deoxycholate and trypsin treatment increased tubular kininogenase activity in an additive fashion. A detailed analysis of microdissected tubule fragments revealed that kallikrein is concentrated in late distal convoluted tubule before entering a branching point (connecting tubule). In contrast initial portions of distal convoluted tubules and cortical collecting tubules contained only little kallikrein activity. Potassium rich diet increased basal and total activity 5-fold, when compared to a potassium poor diet.
使用来自犬血浆的激肽原和缓激肽放射免疫分析法,在显微解剖的小鼠肾单位节段中测量了激肽释放酶的激肽原酶活性。当检查单个肾单位节段时,结果显示出很大的离散度。发现这是由于不同肾单位的远曲小管(DCT)存在异质性,因为在DCT结构池中进行重复测量时并未显示出这种程度的变化。当将新鲜解剖的节段在等渗培养基中孵育时,近20%的活性可被细胞外底物利用。显著释放细胞内酶活性的冻融处理,并未显著提高激肽原酶活性。另一方面,脱氧胆酸盐和胰蛋白酶处理以累加方式增加了肾小管激肽原酶活性。对显微解剖的肾小管片段进行的详细分析表明,激肽释放酶在进入分支点(连接小管)之前集中在远曲小管的后期。相比之下,远曲小管的起始部分和皮质集合小管仅含有很少的激肽释放酶活性。与低钾饮食相比,高钾饮食使基础活性和总活性增加了5倍。