Proteomic expression profiling of Haemophilus influenzae grown in pooled human sputum from adults with chronic obstructive pulmonary disease reveal antioxidant and stress responses.
Proteomic expression profiling of Haemophilus influenzae grown in pooled human sputum from adults with chronic obstructive pulmonary disease reveal antioxidant and stress responses.
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DOI：
10.1186/1471-2180-10-162
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发表时间：
2010-06-01
期刊：
影响因子：
4.2
通讯作者：
Murphy TF
中科院分区：
文献类型：
--
作者：
Qu J;Lesse AJ;Brauer AL;Cao J;Gill SR;Murphy TF
不可分型流感嗜血杆菌定植并感染患有慢性阻塞性肺病的成年人的呼吸道，慢性阻塞性肺疾病是全球第四大常见死亡原因。因此，流感嗜血杆菌是一种人类独有的病原体，已经适应了在人类呼吸道的恶劣环境中生存。为了表征流感嗜血杆菌在呼吸道中表达的蛋白质，在混合的人类痰液中培养原型菌株以模拟人类呼吸道的条件。来自全细菌细胞裂解物的蛋白质被溶解使用强缓冲液，然后使用优化的沉淀/颗粒上酶消化程序进行定量清洁。通过纳流液相色谱/质谱法完成蛋白质组分析，具有低空隙体积和高分离效率以及浅、长梯度。总共鉴定出 1402 个蛋白质，其中包括 170 个由被注释为保守假设蛋白质的基因编码的蛋白质。与化学成分确定的培养基相比，在痰生长条件下，有 31 个蛋白质的丰度更高，比例 > 1.5。其中包括 8 种抗氧化剂和 5 种应激相关蛋白，表明抗氧化活性和应激反应的表达对于气道中的生存很重要。四种蛋白质参与摄取二价阴离子和 9 种具有摄取各种分子功能的蛋白质在痰培养条件下含量更高。与培养基培养的细胞相比，在混合的人痰中生长的流感嗜血杆菌的蛋白质组表达谱显示，抗氧化剂、应激反应蛋白、辅因子和营养吸收系统的表达增加。这些观察结果表明，流感嗜血杆菌通过增加在这些条件下生存所需的分子表达来适应慢性阻塞性肺病成人气道的氧化和营养有限的条件。
Nontypeable Haemophilus influenzae colonizes and infects the airways of adults with chronic obstructive pulmonary disease, the fourth most common cause of death worldwide.Thus, H. influenzae, an exclusively human pathogen, has adapted to survive in the hostile environment of the human airways.To characterize proteins expressed by H. influenzae in the airways, a prototype strain was grown in pooled human sputum to simulate conditions in the human respiratory tract.The proteins from whole bacterial cell lysates were solubilized with a strong buffer and then quantitatively cleaned with an optimized precipitation/on-pellet enzymatic digestion procedure.Proteomic profiling was accomplished by Nano-flow liquid chromatography/mass spectroscopy with low void volume and high separation efficiency with a shallow, long gradient. A total of 1402 proteins were identified with high confidence, including 170 proteins that were encoded by genes that are annotated as conserved hypothetical proteins.Thirty-one proteins were present in greater abundance in sputum-grown conditions at a ratio of > 1.5 compared to chemically defined media.These included 8 anti-oxidant and 5 stress-related proteins, suggesting that expression of antioxidant activity and stress responses is important for survival in the airways.Four proteins involved in uptake of divalent anions and 9 proteins that function in uptake of various molecules were present in greater abundance in sputum-grown conditions. Proteomic expression profiling of H. influenzae grown in pooled human sputum revealed increased expression of antioxidant, stress-response proteins and cofactor and nutrient uptake systems compared to media grown cells.These observations suggest that H. influenzae adapts to the oxidative and nutritionally limited conditions of the airways in adults with chronic obstructive pulmonary disease by increasing expression of molecules necessary for survival in these conditions.
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