Moch T, Hoschützky H, Hacker J, Kröncke K D, Jann K
Proc Natl Acad Sci U S A. 1987 May;84(10):3462-6. doi: 10.1073/pnas.84.10.3462.
The alpha-sialyl-beta-2,3-galactosyl-specific adhesin (S adhesin) was isolated from cells of a recombinant Escherichia coli K-12 strain expressing the S-fimbrial adhesin complex. A crude cell extract was partially dissociated into fimbriae and an adhesin-enriched fraction by heating to 70 degrees C. From the latter, adhesin was purified to apparent homogeneity (by fast protein liquid chromatography, immunoblot, and NaDodSO4/PAGE) by differential ammonium sulfate precipitation, dissociation in 8 M guanidine hydrochloride, and high-resolution anion-exchange chromatography in 8 M urea. The purified adhesin formed an aggregate of Mr approximately 10(6) that was made up of one type of 12-kDa polypeptide (fimbrillin is 16.5 kDa). It had pI value of 4.7 (fimbriae has a pI value of 6). Adhesin and fimbrillin had different amino acid compositions. The purified adhesins agglutinated human and bovine erythrocytes with the same specificity as the whole bacteria; purified fimbriae were not adhesive. Monoclonal anti-adhesin and anti-fimbriae antibodies were obtained. Monoclonal anti-adhesin, but none of the anti-fimbriae, antibodies inhibited the agglutination of erythrocytes. The anti-adhesive antibodies were used in immuno-gold electron microscopy to localize adhesin exclusively on the fimbriae, with a possible preference to their tips.
α-唾液酸-β-2,3-半乳糖基特异性黏附素（S黏附素）是从表达S菌毛黏附素复合物的重组大肠杆菌K-12菌株的细胞中分离得到的。通过加热至70℃，粗细胞提取物部分解离为菌毛和富含黏附素的部分。从后者中，通过硫酸铵分级沉淀、在8M盐酸胍中解离以及在8M尿素中进行高分辨率阴离子交换色谱，将黏附素纯化至表观均一（通过快速蛋白质液相色谱、免疫印迹和十二烷基硫酸钠/聚丙烯酰胺凝胶电泳）。纯化的黏附素形成了一个Mr约为10(6)的聚集体，该聚集体由一种12-kDa的多肽组成（菌毛蛋白为16.5 kDa）。其pI值为4.7（菌毛的pI值为6）。黏附素和菌毛蛋白具有不同的氨基酸组成。纯化的黏附素能以与完整细菌相同的特异性凝集人和牛的红细胞；纯化的菌毛没有黏附性。获得了单克隆抗黏附素和抗菌毛抗体。单克隆抗黏附素抗体，但没有抗菌毛抗体，能抑制红细胞的凝集。抗黏附抗体用于免疫金电子显微镜，将黏附素专门定位在菌毛上，可能优先定位在菌毛尖端。
