Minnick M F, Heinzen R A, Reschke D K, Frazier M E, Mallavia L P
Department of Microbiology, Washington State University, Pullman 99164-4233.
Infect Immun. 1991 Dec;59(12):4735-9. doi: 10.1128/iai.59.12.4735-4739.1991.
The cbbE' gene codes for the E' protein of Coxiella burnetii and was detected in genomic DNA from all known human isolates of the biotzere strain but not in DNA from the other five strains of C. burnetti. The biotzere strain is strictly associated with chronic disease in humans. Extrinsic iodination of biotzere strain cells radiolabeled a 55-kDa protein which comigrated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with the E' protein synthesized in vitro from recombinants containing the cbbE' gene. The 125I-labeled 55-kDa protein was immunoprecipitated with polyclonal anti-E' antiserum, confirming its identity as E'. Predicted secondary structure of the E' polypeptide shows six regions of beta-sheet structure and an alpha-helix near the C terminus with adequate lengths to span a membrane. The predicted hydropathy profile of E' is similar to profiles of known outer membrane proteins and corroborates the biochemical data, indicating that the protein is located in the outer membrane of C. burnetii.
cbbE'基因编码伯氏考克斯氏体的E'蛋白,在生物战剂菌株的所有已知人类分离株的基因组DNA中均可检测到,但在其他五种伯氏考克斯氏体菌株的DNA中未检测到。生物战剂菌株与人类的慢性疾病密切相关。用放射性碘对生物战剂菌株细胞进行体外碘化标记,得到一种55 kDa的蛋白,该蛋白在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳上的迁移情况与含有cbbE'基因的重组体体外合成的E'蛋白相同。用多克隆抗E'抗血清对125I标记的55 kDa蛋白进行免疫沉淀,证实其为E'蛋白。E'多肽的预测二级结构显示有六个β-折叠结构区域,靠近C端有一个α-螺旋,其长度足以跨越细胞膜。E'的预测亲水性图谱与已知外膜蛋白的图谱相似,这与生化数据相符,表明该蛋白位于伯氏考克斯氏体的外膜中。