Paralogous chemoreceptors mediate chemotaxis towards protein amino acids and the non‐protein amino acid gamma‐aminobutyrate (<scp>GABA</scp>)

Rico-Jiménez, Míriam; Muñoz-Martínez, Francisco; García-Fontana, Cristina; Fernández, Matilde; Morel, Bertrand; Ortega, Álvaro; Ramos, Juan L.; Krell, Tino

doi:10.1111/mmi.12255

Cited by 87 publications

(143 citation statements)

References 50 publications

Supporting

Mentioning

134

Contrasting

Order By: Relevance

“…Initially, control experiments were conducted to assess chemotaxis of the wt and mutant strain toward casamino acids. The choice of this chemoattractant was based on the fact that the chemotaxis toward amino acids is mediated by the three well-characterized chemoreceptors PctA, PctB and PctC (Taguchi et al, 1997; Rico-Jimenez et al, 2013). As shown in Supplementary Figure S3 , deletion of mcpK did not alter significantly chemotaxis to casamino acids, indicating that this mutation did not cause any general motility defect.…”

Section: Resultsmentioning

confidence: 99%

“…Examples of the former group are receptors for different L -amino acids (Taguchi et al, 1997; Glekas et al, 2012; Oku et al, 2012; Brennan et al, 2013; Rico-Jimenez et al, 2013; Webb et al, 2016), cyclic organic acids (Luu et al, 2015), purines (Fernandez et al, 2016), polyamines (Corral-Lugo et al, 2016), aromatic hydrocarbons (Lacal et al, 2011b), C4–C6 organic acids (Lacal et al, 2010a; Parales et al, 2013) or C2- and C3-organic acids (Garcia et al, 2015). Chemoreceptors that appear to respond to a single compound include the citrate specific chemoreceptors Tcp of Salmonella typhimurium (Yamamoto and Imae, 1993) and McpQ of P. putida KT2440 (Martin-Mora et al, 2016), the malate specific receptor PA2652 in P. aeruginosa or the GABA specific McpG of P. putida KT2440 (Reyes-Darias et al, 2015a).…”

Section: Discussionmentioning

confidence: 99%

“…In contrast to the dCACHE domain that recognizes ligands in the monomeric state (Rico-Jimenez et al, 2013), the HBM and 4-helix bundle domain need to be dimeric for ligand recognition (Milburn et al, 1991; Lacal et al, 2010a; Martin-Mora et al, 2016). This is due to the fact that the ligand binding sites are at the dimer interface and that amino acids from both monomers of the dimer are involved in ligand binding (Milburn et al, 1991; Pineda-Molina et al, 2012).…”

Section: Discussionmentioning

confidence: 99%

“…In particular its responses to amino acids by the three paralogous receptors PctA, PctB, and PctC (Kuroda et al, 1995; Taguchi et al, 1997; Rico-Jimenez et al, 2013; McKellar et al, 2015; Reyes-Darias et al, 2015a,b) as well as its response to inorganic phosphate (Wu et al, 2000; Rico-Jimenez et al, 2016) by the CtpL and CtpH receptors have been studied in some depth. In addition, the cytosolic and atypical receptor McpB (also named Aer2) (Watts et al, 2011; Airola et al, 2013; Garcia-Fontana et al, 2014; Garcia et al, 2016) was subject to many studies and is amongst the best studied members of the chemoreceptor sub-family with cytoplasmic location.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Identification of a Chemoreceptor in Pseudomonas aeruginosa That Specifically Mediates Chemotaxis Toward α-Ketoglutarate

et al. 2016

Self Cite

View full text Add to dashboard Cite

Pseudomonas aeruginosa is an ubiquitous pathogen able to infect humans, animals, and plants. Chemotaxis was found to be associated with the virulence of this and other pathogens. Although established as a model for chemotaxis research, the majority of the 26 P. aeruginosa chemoreceptors remain functionally un-annotated. We report here the identification of PA5072 (named McpK) as chemoreceptor for α-ketoglutarate (αKG). High-throughput thermal shift assays and isothermal titration calorimetry studies (ITC) of the recombinant McpK ligand binding domain (LBD) showed that it recognizes exclusively α-ketoglutarate. The ITC analysis indicated that the ligand bound with positive cooperativity (Kd1 = 301 μM, Kd2 = 81 μM). McpK is predicted to possess a helical bimodular (HBM) type of LBD and this and other studies suggest that this domain type may be associated with the recognition of organic acids. Analytical ultracentrifugation (AUC) studies revealed that McpK-LBD is present in monomer-dimer equilibrium. Alpha-KG binding stabilized the dimer and dimer self-dissociation constants of 55 μM and 5.9 μM were derived for ligand-free and αKG-bound forms of McpK-LBD, respectively. Ligand-induced LBD dimer stabilization has been observed for other HBM domain containing receptors and may correspond to a general mechanism of this protein family. Quantitative capillary chemotaxis assays demonstrated that P. aeruginosa showed chemotaxis to a broad range of αKG concentrations with maximal responses at 500 μM. Deletion of the mcpK gene reduced chemotaxis over the entire concentration range to close to background levels and wild type like chemotaxis was recovered following complementation. Real-time PCR studies indicated that the presence of αKG does not modulate mcpK expression. Since αKG is present in plant root exudates it was investigated whether the deletion of mcpK altered maize root colonization. However, no significant changes with respect to the wild type strain were observed. The existence of a chemoreceptor specific for αKG may be due to its central metabolic role as well as to its function as signaling molecule. This work expands the range of known chemoreceptor types and underlines the important physiological role of chemotaxis toward tricarboxylic acid cycle intermediates.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Identification of a Chemoreceptor in Pseudomonas aeruginosa That Specifically Mediates Chemotaxis Toward α-Ketoglutarate

et al. 2016

Self Cite

View full text Add to dashboard Cite

show abstract

“…Using isothermal titration calorimetry we have shown that PctA-LBR and PctB-LBR recognize 17 and five l-amino acids, respectively (Rico-Jimé nez et al, 2013). From these experiments we concluded that a good concentration to obtain an excess of ligand for cocrystallization experiments was 3 mM.…”

Section: Resultsmentioning

confidence: 96%

Purification, crystallization and preliminary crystallographic analysis of the ligand-binding regions of the PctA and PctB chemoreceptors fromPseudomonas aeruginosain complex with amino acids

Rico-Jiménez¹,

Muñoz-Martínez²,

Krell³

et al. 2013

Acta Cryst Sect F

Self Cite

View full text Add to dashboard Cite

Pseudomonas aeruginosa is an opportunistic pathogen and one of the major model organisms for the study of chemotaxis. The bacterium harbours 26 genes encoding chemoreceptors, most of which have not been annotated with a function. The paralogous chemoreceptors PctA and PctB (Pseudomonas chemotactic transducer A and B) were found to mediate chemotaxis towards L-amino acids. However, the ligand spectrum of the receptors is quite different since the recombinant ligand-binding region (LBR) of PctA binds 17 different L-amino acids whereas that of PctB recognizes only five. To determine the molecular basis underlying this ligand specificity, PctA-LBR and PctB-LBR have been purified and crystals have been produced after pre-incubation with L-Ile and L-Arg, respectively. Initial crystallization conditions have been identified by the counter-diffusion method and X-ray data have been collected at 2.5 Å (PctA-LBR bound to L-Ile) and 3.14 Å (PctB-LBR bound to L-Arg) resolution. Crystals belonged to space groups P2(1)2(1)2(1) and P3(1)2(1), with unit-cell parameters a = 72.2, b = 78.5, c = 116.6 Å and a = b = 111.6, c = 117.4, respectively, for PctA-LBR and PctB-LBR. Molecular-replacement methods will be pursued for structural determination.

show abstract

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

Cerna‐Vargas,

Krell

2024

MicrobiologyOpen

Self Cite

View full text Add to dashboard Cite

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.

show abstract

Paralogous chemoreceptors mediate chemotaxis towards protein amino acids and the non‐protein amino acid gamma‐aminobutyrate (GABA)

Cited by 87 publications

References 50 publications

Identification of a Chemoreceptor in Pseudomonas aeruginosa That Specifically Mediates Chemotaxis Toward α-Ketoglutarate

Identification of a Chemoreceptor in Pseudomonas aeruginosa That Specifically Mediates Chemotaxis Toward α-Ketoglutarate

Purification, crystallization and preliminary crystallographic analysis of the ligand-binding regions of the PctA and PctB chemoreceptors fromPseudomonas aeruginosain complex with amino acids

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

Contact Info

Product

Resources

About