BIOLOGICAL-ACTIVITIES AND CHEMICAL-COMPOSITION OF PURIFIED TRACHEAL CYTO-TOXIN OF BORDETELLA-PERTUSSIS
BIOLOGICAL-ACTIVITIES AND CHEMICAL-COMPOSITION OF PURIFIED TRACHEAL CYTO-TOXIN OF BORDETELLA-PERTUSSIS
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DOI：
10.1128/iai.57.7.2223-2229.1989
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发表时间：
1989-07-01
影响因子：
3.1
通讯作者：
GOLDMAN, WE
中科院分区：
文献类型：
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作者：
COOKSON, BT;CHO, HL;GOLDMAN, WE
大气道粘膜纤毛上皮细胞的特异性破坏是与人类百日咳杆菌感染相关的原发性呼吸道细胞病理学。我们纯化了一个单一的低分子量糖肽，气管细胞毒素（TCT），它似乎引起了这种病理。采用固相萃取和反相高压液相色谱相结合的方法，从1升百日咳杆菌培养上清液中可分离出约700 nmol的生物活性肽（产率约60%）。浓度为1mu的TCT。M在体外与呼吸道上皮培养时破坏纤毛细胞群。这种TCT浓度与生长中的百日咳杆菌培养上清液中的浓度相似。纯化的TCT还以定量、剂量依赖的方式抑制仓鼠气管上皮细胞的DNA合成。纯化物中未检出内毒素，百日咳和大肠杆菌内毒素均不能复制TCT的生物活性。对纯化后的TCT进行氨基酸和氨基糖分析，发现葡萄糖胺、谷氨酸、丙氨酸和二氨基苯甲酸的摩尔比为1:1:2:1:1。这表明百日咳咳杆菌释放的抑纤毛素TCT是肽聚糖的双糖四肽亚基。
Specific destruction of ciliated epithelial cells lining the large airways is the primary respiratory tract cytopathology associated with human Bordetella pertussis infections. We have purified a single low-molecular-weight glycopeptide, tracheal cytotoxin (TCT), that appears to cause this pathology. By using a combination of solid-phase extraction and reversed-phase high-pressure liquid chromatography, about 700 nmol of biologically active peptide can be isolated from 1 liter of B. pertusis culture supernatant (approximately 60% yield). TCT at concentrations of 1 .mu.M destroyed the ciliated cell population when incubated with respiratory epithelium in vitro. This concentration of TCT is similar to the concentrations found in the culture supernatant of growing B. pertussis. Purified TCT also inhibited DNA synthesis of hamster trachea epithelial cells in a quantitative, dose-dependent fashion. Endotoxin was not detected in the purified material, and neither B. pertussis nor Escherichia coli endotoxin could duplicate the biological activities of TCT. Amino acid and amino sugar analyses of purified TCT revealed the presence of glucosamine, muramic acid, alanine, glutamic acid, and diaminopimelic acid in molar ratios of 1:1:2:1:1. This suggests that TCT, the released ciliostatic principle of B. pertussis, is a disaccharide tetrapeptide subunit of peptidoglycan.