Combining two optimized and affordable methods to assign chemoreceptors to a specific signal

Boyeldieu, Anne; Chaouche, Amine Ali; Méjean, Vincent; Jourlin-Castelli, Cécile

doi:10.1016/j.ab.2021.114139

Cited by 14 publications

(22 citation statements)

References 41 publications

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“…We have previously shown that the chemoreceptor SO_1056, which belongs to the 40H family, is involved in the attraction of S. oneidensis towards chromate [8]. The LBD of SO_1056 contains a double Cache domain (dCache_1).…”

Section: Resultsmentioning

confidence: 99%

“…The recombinant MBP-tagged protein (MBP-LBD1056) was produced from E. coli C600 strain containing the pMAL-LBD-1056 plasmid as previously described [8]. Briefly, the strain was grown aerobically in LB medium at 37 °C until an OD 600 of 0.5 was reached.…”

Section: Expression and Purification Of Mbp-lbd1056 Chimeric Proteinmentioning

confidence: 99%

“…Experimental data are, therefore, necessary to increase our knowledge of signal detection by chemoreceptors. Various approaches are used to couple signal(s) to MCP(s), such as in vivo monitoring of the chemotactic behaviour of mcp-deleted or mcp-overexpressing strains, and in vitro binding assays using purified LBD recombinant proteins (including thermal shift assay, differential scanning fluorimetry and isothermal titration calorimetry) [3,[8][9][10][11].…”

Section: Introductionmentioning

confidence: 99%

“…Seven Cache-containing chemoreceptors were tested for their involvement in chromate attraction. Among them, two allowed an enhanced response towards chromate when overproduced (SO_0987 and SO_1056) but only SO_1056 was able to detect chromate by direct binding to its LBD [8]. This latter chemoreceptor was the focus of this study.…”

Section: Introductionmentioning

confidence: 99%

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Multiple detection of both attractants and repellents by the dCache‐chemoreceptor SO_1056 of Shewanella oneidensis

et al. 2022

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Chemoreceptors are usually transmembrane proteins dedicated to the detection of compound gradients or signals in the surroundings of a bacterium. After detection, they modulate the activation of CheA-CheY, the core of the chemotactic pathway, to allow cells to move upwards or downwards depending on whether the signal is an attractant or a repellent, respectively. Environmental bacteria such as Shewanella oneidensis harbour dozens of chemoreceptors or MCPs (methyl-accepting chemotaxis proteins). A recent study revealed that MCP SO_1056 of S. oneidensis binds chromate. Here, we show that this MCP also detects an additional attractant (L-malate) and two repellents (nickel and cobalt). The experiments were performed in vivo by the agarose-in-plug technique after overproducing MCP SO_1056 and in vitro, when possible, by submitting the purified ligand-binding domain (LBD) of SO_1056 to a thermal shift assay (TSA) coupled to isothermal titration calorimetry (ITC). ITC assays revealed a K D of 3.4 lM for L-malate and of 47.7 lM for nickel. We conclude that MCP SO_1056 binds attractants and repellents of unrelated composition. The LBD of SO_1056 belongs to the double Cache_1 family and is highly homologous to PctA, a chemoreceptor from Pseudomonas aeruginosa that detects several amino acids. Therefore, LBDs of the same family can bind diverse compounds, confirming that experimental approaches are required to define accurate LBD-binding molecules or signals.

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Section: Resultsmentioning

confidence: 99%

Section: Expression and Purification Of Mbp-lbd1056 Chimeric Proteinmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Multiple detection of both attractants and repellents by the dCache‐chemoreceptor SO_1056 of Shewanella oneidensis

et al. 2022

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“…Thermo shift assay was carried out as described previously [ 44 , 45 ]. In brief, each purified protein was mixed with or without 500 μM formic acid in the presence of SYPRO Orange (Thermo Fisher Scientific, Waltham, MA, USA), which was buffered by 50 mM Tris-HCl (pH 7.4).…”

Section: Methodsmentioning

confidence: 99%

Agrobacterium fabrum atu0526-Encoding Protein Is the Only Chemoreceptor That Regulates Chemoattraction toward the Broad Antibacterial Agent Formic Acid

Wang

Zhang

et al. 2021

Biology

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Soil-born plant pathogens, especially Agrobacterium, generally navigate their way to hosts through recognition of the root exudates by chemoreceptors. However, there is still a lack of appropriate identification of chemoreceptors and their ligands in Agrobacterium. Here, Atu0526, a sCache-type chemoreceptor from Agrobacterium fabrum C58, was confirmed as the receptor of a broad antibacterial agent, formic acid. The binding of formic acid to Atu0526 was screened using a thermo shift assay and verified using isothermal titration calorimetry. Inconsistent with the previously reported antimicrobial properties, formic acid was confirmed to be a chemoattractant to A. fabrum and could promote its growth. The chemotaxis of A. fabrum C58 toward formic acid was completely lost with the knock-out of atu0526, and regained with the complementation of the gene, indicating that Atu0526 is the only chemoreceptor for formic acid in A. fabrum C58. The affinity of formic acid to Atu0526LBD significantly increased after the arginine at position 115 was replaced by alanine. However, in vivo experiments showed that the R115A mutation fully abolished the chemotaxis of A. fabrum toward formic acid. Molecular docking based on a predicted 3D structure of Atu0526 suggested that the arginine may provide “an anchorage” for formic acid to pull the minor loop, thereby forming a conformational change that generates the ligand-binding signal. Collectively, our findings will promote an understanding of sCache-type chemoreceptors and their signal transduction mechanism.

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Unveiling preferred chemoattractants for rhizosphere PGPR colonization by molecular docking and molecular dynamics simulations

Chai,

Li,

Gao

et al. 2024

Computers and Electronics in Agriculture

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Combining two optimized and affordable methods to assign chemoreceptors to a specific signal

Cited by 14 publications

References 41 publications

Multiple detection of both attractants and repellents by the dCache‐chemoreceptor SO_1056 of Shewanella oneidensis

Multiple detection of both attractants and repellents by the dCache‐chemoreceptor SO_1056 of Shewanella oneidensis

Agrobacterium fabrum atu0526-Encoding Protein Is the Only Chemoreceptor That Regulates Chemoattraction toward the Broad Antibacterial Agent Formic Acid

Unveiling preferred chemoattractants for rhizosphere PGPR colonization by molecular docking and molecular dynamics simulations

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