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首页> 外文期刊>Journal of bacteriology >Visualization of the Attachment Organelle and Cytadherence Proteins of Mycoplasma pneumoniae by Immunofluorescence Microscopy
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Visualization of the Attachment Organelle and Cytadherence Proteins of Mycoplasma pneumoniae by Immunofluorescence Microscopy

机译：免疫荧光显微镜观察肺炎支原体的附着细胞器和细胞粘附蛋白

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A method was developed for protein localization in Mycoplasma pneumoniae by immunofluorescence microscopy. The P1 adhesin protein was revealed to be located at least at one cell pole in all adhesive cells, as has been observed by immunoelectron microscopy. Cell images were classified according to P1 localization and assigned by DNA content. Cells with a single P1 focus at one cell pole had a lower DNA content than cells with two foci, at least one of which was positioned at a cell pole. Those with one focus at each cell pole had the highest DNA content, suggesting that the nascent attachment organelle is formed next to the old one and migrates to the opposite cell pole before cell division. Double staining revealed that the accessory proteins for cytadherence—HMW1, HMW3, P30, P90, P40, and P65—colocalized with the P1 adhesin in all cells. The localization of cytadherence proteins was also examined in cytadherence-deficient mutant cells with a branched morphology. In M5 mutant cells, which lack the P90 and P40 proteins, HMW1, HMW3, P1, and P30 were focused at the cell poles of short branches, and P65 showed no signal. In M7 mutant cells, which produce a truncated P30 protein, HMW1, HMW3, P1, P90, and P40 were focused, and P65 showed no signal. In M6 mutant cells, which express no HMW1 and a truncated P30 protein, the P1 adhesin was distributed throughout the entire cell body, and no signal was detected for the other proteins. These results suggest that the cytadherence proteins are sequentially assembled to the attachment organelle with HMW1 first, HMW3, P1, P30, P90, and P40 next, and P65 last.
机译：通过免疫荧光显微镜研究了一种肺炎支原体中蛋白质定位的方法。正如免疫电子显微镜所观察到的，P1粘附素蛋白被发现至少位于所有粘附细胞的一个细胞极处。细胞图像根据P1定位进行分类，并按DNA含量分配。一个P1焦点位于一个细胞极的细胞的DNA含量要低于具有两个焦点的细胞，其中至少有一个位于细胞极。那些专注于每个细胞极的细胞具有最高的DNA含量，这表明新生的附着细胞器形成在旧细胞的旁边，并在细胞分裂之前迁移到相反的细胞极。双重染色显示，在所有细胞中，用于细胞粘附的辅助蛋白HMW1，HMW3，P30，P90，P40和P65与P1粘附素共定位。还在具有分支形态的缺乏胞粘附力的突变细胞中检查了胞粘附蛋白的定位。在缺少P90和P40蛋白的M5突变细胞中，HMW1，HMW3，P1和P30集中在短分支的细胞极，而P65没有信号。在产生截短的P30蛋白的M7突变细胞中，对HMW1，HMW3，P1，P90和P40进行了聚焦，而P65未显示信号。在不表达HMW1和截短的P30蛋白的M6突变细胞中，P1粘附素分布在整个细胞体内，没有检测到其他蛋白的信号。这些结果表明，将胞粘附蛋白依次组装至附着细胞器，首先是HMW1，然后是HMW3，P1，P30，P90和P40，最后是P65。

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