Goso Y, Ishihara K, Kurihara M, Sugaya T, Hotta K
Department of Biochemistry, Kitasato University School of Medicine, Kanto Chemical Co., Inc., Isehara, 259-1100, Japan.
J Biochem. 1999 Aug;126(2):375-81. doi: 10.1093/oxfordjournals.jbchem.a022460.
Whole mucins and reduced subunits were extracted from the corpus of the rat stomach. After purification by Sepharose CL-4B chromatography followed by cesium trifluoroacetate equilibrium centrifugation, they were analyzed by Sepharose CL-2B chromatography, rate-zonal sedimentation centrifugation, and Q-Sepharose chromatography. Monoclonal antibodies RGM21 and HIK1083, which histochemically stained mucins in the surface and glandular mucosa of the rat stomach, respectively, were used to detect the site-specific mucins. Although RGM21- and HIK1083-reactive mucins both had a multimerized structure, the density and size of both the whole mucins and reduced subunits differed, thus indicating the presence of distinct mucin species in the surface and glandular mucosa. The mucin subunits were separated into four fractions, UB, B1, B2a, and B2b, by Q-Sepharose chromatography. HIK1083 reacted mainly with UB, while RGM21 reacted with B1, B2a, and B2b. These results, combined with dot-blot, amino acid, and carbohydrate composition analyses, showed that the surface mucins may consist of three kinds of subunits. In contrast, the glandular mucins may consist of one kind of subunit which differs from that of surface mucins.
从大鼠胃体中提取全黏蛋白和还原亚基。经琼脂糖CL - 4B柱层析纯化，再经三氟乙酸铯平衡离心后，通过琼脂糖CL - 2B柱层析、速率区带沉降离心和Q - 琼脂糖柱层析进行分析。分别能对大鼠胃表面和腺黏膜黏蛋白进行组织化学染色的单克隆抗体RGM21和HIK1083，用于检测位点特异性黏蛋白。尽管RGM21和HIK1083反应性黏蛋白均具有多聚化结构，但全黏蛋白和还原亚基的密度和大小不同，这表明在表面和腺黏膜中存在不同种类的黏蛋白。通过Q - 琼脂糖柱层析将黏蛋白亚基分离为四个组分，即UB、B1、B2a和B2b。HIK1083主要与UB反应，而RGM21与B1、B2a和B2b反应。这些结果与斑点印迹、氨基酸和碳水化合物组成分析相结合，表明表面黏蛋白可能由三种亚基组成。相比之下，腺黏蛋白可能由一种与表面黏蛋白不同的亚基组成。
