Monder C, Lakshmi V
Population Council, New York, New York 10021.
Endocrinology. 1990 May;126(5):2435-43. doi: 10.1210/endo-126-5-2435.
Monospecific polyclonal antibodies to purified homogeneous rat liver corticosteroid 11 beta-dehydrogenase were generated in rabbits. The antibodies were immunoprecipitins, but enzyme activity was not completely suppressed in the antigen-antibody complex. Two antibody preparations, 56-125 and 56-126, used to detect 11 beta-dehydrogenase antigen in Western blots, generated different staining patterns for kidney, liver, brain, and heart. Using the two antibodies together, the total number of antibody-reacting components in kidney was three, and that in liver was two. Based on rates of digestion with proteases, the two prominent immunoreactive proteins in kidney appeared to be structurally or conformationally different. A prominent immunostaining component was present in stomach. Tissues that showed immunochemical evidence of 11 beta-dehydrogenase antigen showed corresponding levels of 11 beta-dehydrogenase activity. Most active were liver, testis, kidney, and lung. Lower levels of activity were found in prostate and epididymis, brain, and reproductive tract. We conclude that 11 beta-dehydrogenase is widely distributed in rat organs and is present at low levels with significant exceptions. The data indicate that 11 beta-dehydrogenase may occur in several enzyme forms, and that the distribution of these forms is to some extent tissue specific.
针对纯化的均质大鼠肝脏皮质类固醇11β-脱氢酶,在兔体内产生了单特异性多克隆抗体。这些抗体是免疫沉淀素,但在抗原-抗体复合物中酶活性并未被完全抑制。用于在蛋白质印迹法中检测11β-脱氢酶抗原的两种抗体制剂56 - 125和56 - 126,在肾脏、肝脏、大脑和心脏中产生了不同的染色模式。将这两种抗体一起使用时,肾脏中与抗体反应的成分总数为三种,而肝脏中为两种。根据蛋白酶消化率,肾脏中两种主要的免疫反应性蛋白在结构或构象上似乎有所不同。胃中存在一种显著的免疫染色成分。显示有11β-脱氢酶抗原免疫化学证据的组织呈现出相应水平的11β-脱氢酶活性。活性最高的是肝脏、睾丸、肾脏和肺。在前列腺、附睾、大脑和生殖道中发现的活性水平较低。我们得出结论,11β-脱氢酶在大鼠器官中广泛分布,且除了显著的例外情况外含量较低。数据表明11β-脱氢酶可能以几种酶形式存在,并且这些形式的分布在一定程度上具有组织特异性。