菌Pseudomonas aeruginosa PhuS近接联体His-209触发了从DNA结合到血红转移的功能上的构造性切换
Nicholas Montes1, Tyree Wilson2, Samuel A Krug1
1Department of Pharmaceutical Sciences, School of Pharmacy, University of Maryland, Baltimore, Maryland, USA.
The Journal of biological chemistry
|March 23, 2025
概括
Pseudomonas aeruginosa 使用 PhuS 蛋白调节从血中吸收铁. 佛山的His-209对于其在血红素转移和基因调节中的双重功能至关重要,影响细菌适应.
科学领域:
- 微生物学 微生物学
- 分子生物学分子生物学
- 生物化学 生物化学
背景情况:
- 伪菌 (Pseudomonas aeruginosa) 使用血红素通过特定的吸收系统来获取铁.
- 细胞质蛋白 PhuS 通过血红氧酶 HemO 调节血红流.
- S表现出双重功能,控制细胞外血红流,并在它的apo形式,调节小RNAs (sRNAs) PrrF/PrrH.
研究的目的:
- 研究His-209在PhuS功能中的作用及其对海姆吸收和sRNA调节的影响.
- 为了阐明突变的结构和功能后果的His-209残留在福州.
- 了解PhuS如何将细胞外血利用集成到PrrF/PrrHsRNA调节网络中.
主要方法:
- 电泳移动转移试验 (EMSA) 和光异性,以评估DNA结合亲和力.
- -交换质谱法 (HDX-MS) 用于分析蛋白质结构.
- 定量PCR (qPCR) 和西部涂抹来评估基因和蛋白质表达.
主要成果:
- 对His-209 (PhuS H209A) 的突变显著降低了apo和holo-PhuS对prrF1促进物的结合亲和力.
- 结构分析显示,apo-和holo-PhuS H209A采用野生类型apo-和holo-PhuS之间的中间形状.
- 斯H209A突变体表现出未合的血红素流量调节,废除了PrrF/PrrHsRNAs的血红素依赖控制.
结论:
- 通过His-209进行血质协调对于决定PhuS双重监管功能的结构转换至关重要.
- 这种His-209残留物对于PhuS在血红素转移和DNA结合中的作用的相互排他性至关重要.
- 在PrrF/PrrHsRNA调节网络中,PhuS作为细胞外血代谢的关键整合剂,对P. aeruginosa宿主适应至关重要.
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