Exploring solute binding proteins in Pseudomonas aeruginosa that bind to γ-aminobutyrate and 5-aminovalerate and their role in activating sensor kinases

IF 3.9 3区生物学 Q2 MICROBIOLOGY

MicrobiologyOpen Pub Date : 2024-05-23 DOI:10.1002/mbo3.1415

Jean Paul Cerna-Vargas, Tino Krell

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Abstract

The standard method of receptor activation involves the binding of signals or signal-loaded solute binding proteins (SBPs) to sensor domains. Many sensor histidine kinases (SHKs), which are activated by SBP binding, are encoded adjacent to their corresponding sbp gene. We examined three SBPs of Pseudomonas aeruginosa PAO1, encoded near the genes for the AgtS (PA0600) and AruS (PA4982) SHKs, to determine how common this arrangement is. Ligand screening and microcalorimetric studies revealed that the SBPs PA0602 and PA4985 preferentially bind to GABA (KD = 2.3 and 0.58 μM, respectively), followed by 5-aminovalerate (KD = 30 and 1.6 μM, respectively) and ethanoldiamine (KD = 2.3 and 0.58 μM, respectively). In contrast, AgtB (PA0604) exclusively recognizes 5-aminovaleric acid (KD = 2.9 μM). However, microcalorimetric titrations did not show any binding between the AgtS sensor domain and AgtB or PA0602, regardless of the presence of ligands. Similarly, bacterial two-hybrid assays did not demonstrate an interaction between PA4985 and the AruS sensor domain. Therefore, sbp and shk genes located nearby are not always functionally linked. We previously identified PA0222 as a GABA-specific SBP. The presence of three SBPs for GABA may be linked to GABA's role as a trigger for P. aeruginosa virulence.

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探索铜绿假单胞菌中与γ-氨基丁酸和 5-氨基戊酸结合的溶质结合蛋白及其在激活感应激酶中的作用。

受体激活的标准方法是将信号或信号负载的溶质结合蛋白（SBP）与传感器结构域结合。许多由 SBP 结合激活的传感器组氨酸激酶（SHKs）的编码与相应的 sbp 基因相邻。我们研究了铜绿假单胞菌 PAO1 的三个 SBPs（在 AgtS（PA0600）和 AruS（PA4982）SHKs 基因附近编码），以确定这种排列的常见程度。配体筛选和微量热研究显示，SBPs PA0602 和 PA4985 优先与 GABA 结合（KD 分别为 2.3 和 0.58 μM），其次是 5-氨基戊酸（KD 分别为 30 和 1.6 μM）和乙醇胺（KD 分别为 2.3 和 0.58 μM）。相比之下，AgtB（PA0604）只能识别 5-氨基戊酸（KD = 2.9 μM）。然而，微量热滴定法并未显示 AgtS 传感器结构域与 AgtB 或 PA0602 有任何结合，无论是否存在配体。同样，细菌双杂交试验也没有证明 PA4985 与 AruS 传感器结构域之间存在相互作用。因此，位于附近的 sbp 和 shk 基因在功能上并不总是相关联的。我们之前发现 PA0222 是一种 GABA 特异性 SBP。GABA 的三个 SBP 的存在可能与 GABA 作为铜绿假单胞菌毒力触发器的作用有关。

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来源期刊

MicrobiologyOpen MICROBIOLOGY-

CiteScore

8.00

自引率

0.00%

发文量

审稿时长

20 weeks

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