Leyh R, Griffith R W
Department of Microbiology, Immunology, and Preventive Medicine, Iowa State University, Ames 50011.
Infect Immun. 1992 Mar;60(3):958-64. doi: 10.1128/iai.60.3.958-964.1992.
The outer membrane proteins of Bordetella avium were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Sarkosyl-insoluble outer membrane protein-enriched profiles from 50 virulent B. avium isolates, containing major 21,000- and 37,000-molecular-weight proteins (21K and 37K proteins, respectively) and at least 13 less intensely stained proteins with molecular weights ranging from 13,500 to 143,000, were very similar. The 21K, 27K, 31K, and 37K outer membrane proteins were shown to be associated noncovalently with the underlying peptidoglycan layer. It was necessary to treat cell envelopes with 2% sodium dodecyl sulfate and at temperatures in excess of 60 degrees C for 15 min to release these proteins. Exposure of proteins on the cell surface of B. avium was assessed by labeling with 125I followed by electrophoresis. As many as 13 bands were present in profiles from labeled whole cells. Of the surface-labeled bands, eight corresponded to bands in a radiolabeled outer membrane preparation. The outer membrane protein profile of B. avium was compared with profiles from other Bordetella spp., including 20 B. avium-like and 16 B. bronchiseptica strains isolated from turkeys. The outer membrane protein profile of B. avium was distinctly different from those of the other bordetella. The effect of variations in the growth medium on the expression of outer membrane proteins of B. avium was examined. Expression of 22K, 26K, 56K, and 73K proteins was decreased or eliminated by addition of 50 mM MgSO4 to the medium.
通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳对禽博德特氏菌的外膜蛋白进行了检测。来自50株禽博德特氏菌强毒株的 Sarkosyl 不溶性外膜蛋白富集图谱非常相似，其中含有主要的分子量为21,000和37,000的蛋白（分别为21K和37K蛋白）以及至少13条染色较浅、分子量在13,500至143,000之间的蛋白条带。结果表明，21K、27K、31K和37K外膜蛋白与下面的肽聚糖层非共价结合。必须用2%的十二烷基硫酸钠在超过60摄氏度的温度下处理细胞包膜15分钟才能释放这些蛋白。通过用125I标记然后进行电泳来评估禽博德特氏菌细胞表面蛋白的暴露情况。来自标记全细胞的图谱中存在多达13条带。在表面标记的条带中，有8条与放射性标记的外膜制剂中的条带相对应。将禽博德特氏菌的外膜蛋白图谱与其他博德特氏菌属的图谱进行了比较，包括从火鸡中分离出的20株类禽博德特氏菌和16株支气管败血博德特氏菌菌株。禽博德特氏菌的外膜蛋白图谱与其他博德特氏菌的图谱明显不同。研究了生长培养基变化对禽博德特氏菌外膜蛋白表达的影响。向培养基中添加50 mM MgSO4会降低或消除22K、26K、56K和73K蛋白的表达。
