Immunolocalization of betaine aldehyde dehydrogenase in porcine kidney
Immunolocalization of betaine aldehyde dehydrogenase in porcine kidney
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DOI：
10.1006/bbrc.1999.0584
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发表时间：
1999-05-19
影响因子：
3.1
通讯作者：
Valenzuela-Soto, EM
中科院分区：
文献类型：
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作者：
Figueroa-Soto, CG;Lopez-Cervantes, G;Valenzuela-Soto, EM
在兔中产生针对从猪肾纯化的天然BADH的多克隆抗BADH血清。该抗血清与肾、苋菜、铜绿假单胞菌的BADH有较强的交叉反应（1：1000），与苋菜的BADH有较弱的交叉反应（1：100）。在免疫印迹中结合纯化的天然肾BADH的抗体显示出表观分子量为340 kDa的主带和表观分子量为52 kDa的亚基。活性测定的数据显示，皮质部分（81.3 nmol/min/mg蛋白质）比髓质部分（21.3 nmol/min/mg蛋白质）的活性更高。肾组织切片中BADH的免疫定位显示BADH存在于皮质和髓质中。在髓质内，酶主要定位于小管周围的细胞。皮质和髓质切片提取物的Western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相一致。(C)北京：科学出版社.
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. (C) 1999 Academic Press.