Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, United States of America.
PLoS One. 2010 Nov 2;5(11):e13800. doi: 10.1371/journal.pone.0013800.
We examined global changes in protein expression in the B31 strain of Borrelia burgdorferi, in response to two environmental cues (pH and temperature) chosen for their reported similarity to those encountered at different stages of the organism's life cycle. Multidimensional nano-liquid chromatographic separations coupled with tandem mass spectrometry were used to examine the array of proteins (i.e., the proteome) of B. burgdorferi for different pH and temperature culture conditions. Changes in pH and temperature elicited in vitro adaptations of this spirochete known to cause Lyme disease and led to alterations in protein expression that are associated with increased microbial pathogenesis. We identified 1,031 proteins that represent 59% of the annotated genome of B. burgdorferi and elucidated a core proteome of 414 proteins that were present in all environmental conditions investigated. Observed changes in protein abundances indicated varied replicon usage, as well as proteome functional distributions between the in vitro cell culture conditions. Surprisingly, the pH and temperature conditions that mimicked B. burgdorferi residing in the gut of a fed tick showed a marked reduction in protein diversity. Additionally, the results provide us with leading candidates for exploring how B. burgdorferi adapts to and is able to survive in a wide variety of environmental conditions and lay a foundation for planned in situ studies of B. burgdorferi isolated from the tick midgut and infected animals.
我们研究了伯氏疏螺旋体 B31 株对两种环境信号（pH 值和温度）的全球蛋白质表达变化，这两种环境信号因其与该生物生命周期不同阶段所经历的环境相似而被选择。多维纳诺液相色谱分离与串联质谱联用技术被用于研究不同 pH 值和温度培养条件下伯氏疏螺旋体的蛋白质组（即蛋白质组）。体外适应实验表明，pH 值和温度的变化可引起这种已知会引起莱姆病的螺旋体的适应，导致与增加微生物发病机制相关的蛋白质表达改变。我们鉴定了 1031 种蛋白质，它们代表了伯氏疏螺旋体注释基因组的 59%，并阐明了在所有研究环境条件下都存在的 414 种核心蛋白质组。观察到的蛋白质丰度变化表明，在体外细胞培养条件下，复制子的使用以及蛋白质组功能分布存在差异。令人惊讶的是，模拟伯氏疏螺旋体在饱食的蜱肠道中生存的 pH 值和温度条件下，蛋白质多样性明显减少。此外，这些结果为我们提供了探索伯氏疏螺旋体如何适应和能够在各种环境条件下生存的主要候选对象，并为计划从蜱肠道和感染动物中分离的伯氏疏螺旋体进行原位研究奠定了基础。
