Identical phosphatase mechanisms achieved through distinct modes of binding phosphoprotein substrate

Pazy, Y.; Motaleb, M. A.; Guarnieri, Michael T.; Charon, Nyles W.; Zhao, Rui; Silversmith, Ruth E.

doi:10.1073/pnas.0911185107

Cited by 44 publications

(77 citation statements)

References 43 publications

(47 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The NarX Q404A substitution abrogated phosphatase function, as did the analogous CheZ Q147A and CheX N99A substitutions (12,14). The NarX Q404E substitution likewise eliminated phosphatase activity, whereas the Q404N mutant exhibited appreciable activity.…”

Section: Discussionmentioning

confidence: 92%

“…1D) (23). The CheX conserved Asn-99 and Glu-96 residues within this motif function in virtually identical manner to CheZ Gln-147 and Asp-143 (14).…”

Section: Discussionmentioning

confidence: 98%

“…In CheZ, the conserved Gln-147 orients the nucleophilic water for an in-line hydrolysis of the phosphoryl group (12). The auxiliary phosphatase CheX employs a virtually identical mechanism with an analogous role for Asn-99 (14). Likewise, Ras-like GTPases rely on a conserved Gln or Asn for the correct alignment of the nucleophilic water in GTP hydrolysis (13).…”

Section: Discussionmentioning

confidence: 99%

“…Both genetic and biochemical data demonstrate the requirement of residues Gln-404 and Asp-400 for NarX transmitter phosphatase activity. Studies with chemotaxis auxiliary phosphatases and Raslike GTPases have revealed that conserved Gln or Asn residues catalyze phosphoryl group hydrolysis by coordinating the nucleophilic water (12)(13)(14). We therefore propose that Gln-404 catalyzes this reaction in NarX and related sensors.…”

mentioning

confidence: 99%

See 3 more Smart Citations

Conserved mechanism for sensor phosphatase control of two-component signaling revealed in the nitrate sensor NarX

Huynh¹,

Noriega

Stewart

2010

Proc. Natl. Acad. Sci. U.S.A.

100

124

View full text Add to dashboard Cite

Two-component signal transduction mediates a wide range of phenotypes in microbes and plants. The sensor transmitter module controls the phosphorylation state of the cognate-responseregulator receiver domain. Whereas the two-component autokinase and phosphotransfer reactions are well-understood, the mechanism by which sensors accelerate the rate of phosphoresponse regulator dephosphorylation, termed "transmitter phosphatase activity," is unknown. We identified a conserved DxxxQ motif adjacent to the phospho-accepting His residue in the HisKA_3 subfamily of two-component sensors. We used site-specific mutagenesis to make substitutions for these conserved Gln and Asp residues in the nitrate-responsive NarX sensor and analyzed function both in vivo and in vitro. Results show that the Gln residue is critical for transmitter phosphatase activity, but is not essential for autokinase or phosphotransfer activities. The documented role of an amide moiety in phosphoryl group hydrolysis suggests an analogous catalytic function for this Gln residue in HisKA_3 members. Results also indicate that the Asp residue is important for both autokinase and transmitter phosphatase activities. Furthermore, we noted that sensors of the HisKA subfamily exhibit an analogous E/DxxT/N motif, the conserved Thr residue of which is critical for transmitter phosphatase activity of the EnvZ sensor. Thus, twocomponent sensors likely use similar mechanisms for receiver domain dephosphorylation.histidine kinase | DHp domain | DesK

show abstract

Section: Discussionmentioning

confidence: 92%

“…1D) (23). The CheX conserved Asn-99 and Glu-96 residues within this motif function in virtually identical manner to CheZ Gln-147 and Asp-143 (14).…”

Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Conserved mechanism for sensor phosphatase control of two-component signaling revealed in the nitrate sensor NarX

Huynh¹,

Noriega

Stewart

2010

Proc. Natl. Acad. Sci. U.S.A.

100

124

View full text Add to dashboard Cite

show abstract

“…1U0S ); 2CH4 , 2F9Z (Chao et al 2006) and 3HZH (Pazy et al 2010). More information on templates can be found in Table 1.…”

Section: Homology Modelling and Validationmentioning

confidence: 99%

Comparative structural bioinformatics analysis of Bacillus amyloliquefaciens chemotaxis proteins within Bacillus subtilis group

Yssel

Reva

Bishop

2011

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

Chemotaxis is a process in which bacteria sense their chemical environment and move towards more favourable conditions. Since plant colonization by bacteria is a multifaceted process which requires a response to the complex chemical environment, a finely tuned and sensitive chemotaxis system is needed. Members of the Bacillus subtilis group including Bacillus amyloliquefaciens are industrially important, for example in bio-pesticides. The group exhibits plant growth promoting characteristics, with different specificity towards certain host plants. Therefore, we hypothesize that while the principal molecular mechanisms of bacterial chemotaxis may be conserved, the bacterial chemotaxis system may need an evolutionary tweaking to adapt it to specific requirements, particularly in the process of evolution of free-living soil organisms, towards plant colonization behaviour. To date almost nothing is known about what parts of the chemotaxis proteins are subjected to positive amino acid substitutions, involved in adjusting the chemotaxis system of bacteria during speciation.In this novel study, positively selected and purified sites of chemotaxis proteins were calculated and these residues were mapped onto homology models that were built for the chemotaxis proteins, in an attempt to understand the spatial evolution of the chemotaxis proteins. Various positively selected amino acids were identified in semi-conserved regions of the proteins away from the known active sites.

show abstract

Protein Dynamics in Phosphoryl-Transfer Signaling Mediated by Two-Component Systems

Trajtenberg

Buschiazzo

2019

Methods in Molecular Biology

View full text Add to dashboard Cite

The ability to perceive the environment, an essential attribute in living organisms, is linked to the evolution of signalling proteins that recognize specific signals and execute predetermined responses. Such proteins constitute concerted systems that can be as simple as a unique protein, able to recognize a ligand and exert a phenotypic change, or extremely complex pathways engaging dozens of different proteins which act in coordination with feedback loops and signal modulation. To understand how cells sense their surroundings and mount specific adaptive responses, we need to decipher the molecular workings of signal recognition, internalization, transfer and conversion into chemical changes inside the cell. Protein allostery and dynamics play a central role. Here, we review recent progress on the study of twocomponent systems, important signalling machineries of prokaryotes and lower eukaryotes.Such systems implicate a sensory histidine-kinase and a separate response regulator protein.Both components exploit protein flexibility to effect specific conformational rearrangements, modulating protein:protein interactions, and ultimately transmitting information accurately.Recent work has revealed how histidine-kinases switch between discrete functional states according to the presence or absence of the signal, shifting key amino acid positions that define their catalytic activity. In concert with the cognate response regulator's allosteric changes, the phosphoryl-transfer flow during the signalling process is exquisitely fine-tuned for proper specificity, efficiency and directionality.

show abstract

Identical phosphatase mechanisms achieved through distinct modes of binding phosphoprotein substrate

Cited by 44 publications

References 43 publications

Conserved mechanism for sensor phosphatase control of two-component signaling revealed in the nitrate sensor NarX

Conserved mechanism for sensor phosphatase control of two-component signaling revealed in the nitrate sensor NarX

Comparative structural bioinformatics analysis of Bacillus amyloliquefaciens chemotaxis proteins within Bacillus subtilis group

Protein Dynamics in Phosphoryl-Transfer Signaling Mediated by Two-Component Systems

Contact Info

Product

Resources

About