Kishi M, Nakajo S, Shibayama T, Nakaya K, Nakamura Y
J Biochem. 1980 Jan;87(1):135-41. doi: 10.1093/oxfordjournals.jbchem.a132718.
The cell surface structure of AH-66 hepatoma ascites cells was examined by extracting the intact AH-66 cells with urea and analyzing the extracted proteins. When AH-66 cells were suspended in 1 M urea, material composed of approximately 90% protein and 10% carbohydrate was released. The extracted proteins amounted to about 3% of the total cell proteins and were composed of approximately 30 species as analyzed by sodium dodecyl sulfate gel electrophoresis. The major bands had apparent molecular weights of 84,000 and 50,000--60,000 on the gel. In marked contrast to chick embryo fibroblasts, the extracted proteins contained no components stainable with periodic acid-Schiff reagent. Lactoperoxidase-catalyzed iodination of intact AH-66 cells showed that most of the urea-extractable proteins were located on the outer surface of the plasma membranes of AH-66 cells. It was also found that there are two integral proteins exposed on the outer surface of the plasma membranes of AH-66 cells; one is a major glycoprotein (with a molecular weight of 165,000) and the other is a periodate-Schiff-negative protein with a molecular weight of 130,000.
通过用尿素提取完整的AH - 66细胞并分析提取的蛋白质，对AH - 66肝癌腹水细胞的细胞表面结构进行了研究。当AH - 66细胞悬浮于1M尿素中时，会释放出由约90%蛋白质和10%碳水化合物组成的物质。提取的蛋白质约占细胞总蛋白的3%，经十二烷基硫酸钠凝胶电泳分析，这些蛋白质由大约30种成分组成。凝胶上主要条带的表观分子量分别为84,000和50,000 - 60,000。与鸡胚成纤维细胞形成显著对比的是，提取的蛋白质中没有能用高碘酸 - 席夫试剂染色的成分。乳过氧化物酶催化的完整AH - 66细胞碘化显示，大多数可被尿素提取的蛋白质位于AH - 66细胞质膜的外表面。还发现AH - 66细胞质膜外表面有两种整合蛋白；一种是主要糖蛋白（分子量为165,000），另一种是分子量为130,000的高碘酸 - 席夫阴性蛋白。
