A domain-specific marker for the hepatocyte plasma membrane: localization of leucine aminopeptidase to the bile canalicular domain.
A domain-specific marker for the hepatocyte plasma membrane: localization of leucine aminopeptidase to the bile canalicular domain.
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DOI：
10.1083/jcb.96.6.1548
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发表时间：
1983-06
期刊：
影响因子：
--
通讯作者：
Hubbard AL
中科院分区：
文献类型：
--
作者：
Roman LM;Hubbard AL
间接免疫荧光法用于建立肝细胞质膜的结构域特异性标记物。在固定大鼠肝脏的冷冻切片（0.5 - 4微米）中，针对大鼠肠亮氨酸氨基肽酶（亮氨酸氨基肽酶）的抗体识别仅限于胆小管质膜的抗原。在窦状隙或侧膜上未观察到荧光，也未检测到细胞内染色。基于其与肠抗原的化学相似性，将肝抗原鉴定为抗-HCV。首先，使用胰蛋白酶溶解的肠内毒素（G-200级分，91%纯度）的免疫沉淀实验建立了可溶级分中的肠内毒素酶活性损失与在SDS PAGE上迁移110，000和130，000道尔顿之间的多肽的特异性免疫沉淀物中的出现之间的相关性。从肝质膜的洗涤剂提取物中沉淀的抗原具有相同的电泳迁移率。第二，在肝脏免疫沉淀物中的主要条带的胰凝乳蛋白酶图谱与纯化的肠上皮细胞相似。
Indirect immunofluorescence was used to establish a domain-specific marker for hepatocyte plasma membranes. In frozen sections of fixed rat liver (0.5-4 microns), antibodies directed against rat intestinal leucine aminopeptidase (LAP) recognized an antigen that was restricted to the bile canalicular plasma membrane. Fluorescence was not observed on the sinusoidal or lateral membranes, and intracellular staining was not detected. The liver antigen was identified as LAP, based on its chemical similarity to intestinal LAP. First, immunoprecipitation experiments using trypsin-solubilized intestinal LAP (G-200 fraction, 91% pure) established a correlation between the loss of LAP enzyme activity from the soluble fraction and the appearance in the specific immunoprecipitates of polypeptides migrating on SDS PAGE between 110,000 and 130,000 daltons. The antigen precipitated from a detergent extract of liver plasma membranes had the same electrophoretic mobility. Second, the chymotryptic map of the major band in the liver immunoprecipitate was similar to that of purified intestinal LAP.