Figueroa-Soto C G, Lopez-Cervantes G, Valenzuela-Soto E M
Dirección de Ciencia de los Alimentos, Centro de Investigación en Alimentación y Desarrollo A.C., Hermosillo, Sonora, México.
Biochem Biophys Res Commun. 1999 May 19;258(3):732-6. doi: 10.1006/bbrc.1999.0584.
Polyclonal anti-BADH serum was raised in rabbits against native BADH purified from porcine kidney. The antiserum cross-reacted strongly with BADH purified from kidney, Amaranthus palmierii, and Pseudomona aeuroginosa (1:1000), and weakly with Amaranthus hypochondriacus L (1:100). Antibodies bound to purified native kidney BADH in immunoblots showed a major band of an apparent molecular mass of 340 kDa and a subunit with an apparent molecular mass of 52 kDa. Data on activity assays showed higher activity in cortex sections (81.3 nmol/min/mg protein) than in medulla sections (21.3 nmol/min/mg protein). Immunolocalization of BADH in kidney tissue sections showed that BADH is found in cortex and medulla. In inner medulla, the enzyme was mainly localized in cells surrounding the tubules. Western blot analysis on extracts from the cortex and medulla sections showed higher concentration of BADH protein in cortex than in medulla. These results were in accordance with immunolocalization and activity analysis.
用从猪肾中纯化的天然BADH免疫家兔制备多克隆抗BADH血清。该抗血清与从肾、皱果苋和铜绿假单胞菌中纯化的BADH发生强烈交叉反应（1:1000），与低地苋的交叉反应较弱（1:100）。免疫印迹中与纯化的天然肾BADH结合的抗体显示出一条表观分子量为340 kDa的主要条带和一个表观分子量为52 kDa的亚基。活性测定数据显示，皮质切片中的活性（81.3 nmol/分钟/毫克蛋白质）高于髓质切片（21.3 nmol/分钟/毫克蛋白质）。肾组织切片中BADH的免疫定位显示，BADH存在于皮质和髓质中。在内髓质中，该酶主要定位于肾小管周围的细胞中。对皮质和髓质切片提取物的蛋白质免疫印迹分析显示，皮质中BADH蛋白的浓度高于髓质。这些结果与免疫定位和活性分析结果一致。
