Rose M C, Brown C F, Jacoby J Z, Lynn W S, Kaufman B
Department of Child Health and Development, George Washington University, Children's Hospital National Medical Center, Washington, D.C. 20010.
Pediatr Res. 1987 Nov;22(5):545-51. doi: 10.1203/00006450-198711000-00015.
Tracheobronchial mucins from healthy individuals and from patients with bronchial asthma or cystic fibrosis (CF) were isolated from lung mucus, purified, and their chemical and physical properties compared. Normal and asthmatic mucins required both a dissociating and a reducing agent for solubilization and exhibited identical chromatographic behavior on Sepharose 4B, Sepharose 2B, and hydroxylapatite and similar amino acid and carbohydrate compositions. In contrast, 1) CF lung mucins were solubilized in the absence of dissociating and/or reducing agents and 2) the majority of the CF mucins analyzed was eluted in the included volume of Sepharose 4B with Kd values of 0.3 +/- 0.1 rather than in the void volume and thus appeared smaller than normal and asthmatic mucins. The lower molecular weight mucins in CF sputum apparently are produced by bacterial or inflammatory cell proteinases since radiolabelled asthmatic mucin was digested to smaller fragments when incubated with crude CF lung mucosal samples. Furthermore, mucins secreted by tracheal explants from CF and from non-CF individuals eluted in the void volume on Sepharose 4B, suggesting that CF tracheobronchial mucins were not inherently smaller than non-CF mucins.
从健康个体以及患有支气管哮喘或囊性纤维化(CF)的患者的肺黏液中分离出气管支气管黏蛋白,进行纯化,并比较其化学和物理性质。正常和哮喘黏蛋白溶解需要解离剂和还原剂,在琼脂糖4B、琼脂糖2B和羟基磷灰石上表现出相同的色谱行为,且氨基酸和碳水化合物组成相似。相比之下,1)CF肺黏蛋白在没有解离剂和/或还原剂的情况下即可溶解,2)分析的大多数CF黏蛋白在琼脂糖4B的内水体积中洗脱,Kd值为0.3±0.1,而不是在空体积中洗脱,因此看起来比正常和哮喘黏蛋白小。CF痰液中较低分子量的黏蛋白显然是由细菌或炎症细胞蛋白酶产生的,因为放射性标记的哮喘黏蛋白与CF肺黏膜粗样品一起孵育时会被消化成较小的片段。此外,CF个体和非CF个体的气管外植体分泌的黏蛋白在琼脂糖4B的空体积中洗脱,这表明CF气管支气管黏蛋白并非天生就比非CF黏蛋白小。