Escherichia coli aspartate receptor. Oligomerization of the cytoplasmic fragment

Long, D.G.; Weis, Robert M.

doi:10.1016/s0006-3495(92)81782-4

Cited by 9 publications

(9 citation statements)

References 11 publications

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“…The Κ Ά value for the dimer formation was estimated to be 10 5 M _1 , which is similar to the K a value for the dimer formation of the cytoplasmic domain of the aspartate receptor, Tar [17,18]. The present results provide insight into the structure and function of the cytoplasmic signaling domain of EnvZ.…”

Section: Introductionsupporting

confidence: 76%

“…However, it appeared to be a monomer protein because it did not show the monomer-dimer equilibrium on the column after dilution of the protein solution. The monomer EnvZ(C) might have a non-globular molecular shape like a cytoplasmic domain of Tar protein [17,18]. Therefore, in order to estimate the Κ Ά value for the dimer formation of EnvZ(C), we developed a binding assay system to determine the oligomer formation using the Ni-histidine tag affinity.…”

Section: Determination Of Apparent Ka Value Of Envz(c) Dimermentioning

confidence: 99%

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Demonstration of dimer formation of the cytoplasmic domain of a transmembrane osmosensor protein, EnvZ, of Escherichia coli using Ni‐histidine tag affinity chromatography

1997

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EnvZ is a transmembrane osmosensor which regulates the phosphorylation of OmpR, a transcription factor for ompF and ompC genes which encode the major outer membrane porin proteins, OmpF and OmpC in Escherichia coli. Autophosphorylation of EnvZ occurs through a transphosphorylation reaction between two EnvZ molecules. To elucidate the molecular mechanism of signal transduction by EnvZ, we examined the dimer formation of the EnvZ cytoplasmic domain [EnvZ(C)]. For this purpose, we developed a method to determine the complex formation between the purified EnvZ(C) and the purified His6-EnvZ(C) by means of Ni-6xhistidine tag affinity chromatography. When the mixture of EnvZ(C) and His6-EnvZ(C) was applied to Ni-NTA resin, both His6-EnvZ(C) and EnvZ(C) were bound to the resin, indicating that EnvZ can form an oligomer without the periplasmic and transmembrane domains. Binding experiments using the Ni-NTA resin revealed that EnvZ(C) forms a dimer with the K a value for dimerization being approximately 10 5 M _1 in the equilibrium state.

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Section: Introductionsupporting

confidence: 76%

Section: Determination Of Apparent Ka Value Of Envz(c) Dimermentioning

confidence: 99%

Demonstration of dimer formation of the cytoplasmic domain of a transmembrane osmosensor protein, EnvZ, of Escherichia coli using Ni‐histidine tag affinity chromatography

1997

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“…In both cases, a qualitatively similar pattern is observed with or without vesicles (gray and black bars, respectively), suggesting that the reactivity differences are due to intrinsic properties of CF monomers (CF in solution has been shown to be monomeric under similar conditions). 17,28 …”

Section: Resultsmentioning

confidence: 99%

Membrane Association of a Protein Increases the Rate, Extent, and Specificity of Chemical Cross-Linking

et al. 2013

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Many cellular processes involve interactions between membrane-associated proteins, and those interactions are enhanced by membrane association. We have used crosslinking reactions to compare the extent and specificity of protein interactions in solution versus on a membrane surface. Cysteine mutants of a soluble cytoplasmic fragment (CF) of the aspartate receptor, a transmembrane receptor involved in bacterial chemotaxis, are used in disulfide bond formation with the thiol-specific oxidant diamide and chemical crosslinking reactions with the trifunctional maleimide TMEA. CF binding to vesicles is mediated by its N-terminal His tag binding to vesicles containing a nickel-chelating lipid, so crosslinking reactions conducted in the presence and absence of vesicles differ only in whether CF is bound to the vesicles or is free in solution. For multiple Cys throughout the CF, membrane association is shown to increase the rate and extent of these reactions. Crosslinking specificity, which is measured as the preference for crosslinking between Cys near each other in the native structure, is also enhanced by membrane association. These results provide an experimental demonstration that membrane binding enhances protein-protein interactions, an important consideration for understanding processes involving membrane-associated proteins. The experiments further demonstrate the importance of crosslinking conditions for these reactions that are often used to probe protein structure and dynamics, and the potential of membrane association to restore native interactions of membrane-associated proteins for crosslinking studies.

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“…The methyltransferase CheR binds tightly to the cytoplasmic domains of the chemotaxis receptors [43,44]. The binding site has recently been localized to a five amino acid sequence, Asn-Trp-Glu-Thr-Phe, located at the extreme C termini of the E. coli and S. typhimurium aspartate receptors (Tar) and the E. coli serine receptor (Tsr).…”

Section: Interaction Between the Methyltransferase And Its Receptor Substratementioning

confidence: 99%

Crystal structure of the chemotaxis receptor methyltransferase CheR suggests a conserved structural motif for binding S-adenosylmethionine

1997

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The CheR structure shares some structural similarities with small molecule DNA and RNA methyltransferases, despite a lack of sequence similarity among them. In particular, there is significant structural preservation of the S-adenosylmethionine-binding clefts; the specific length and conformation of a loop in the alpha/beta domain seems to be required for S-adenosylmethionine binding within these enzymes. Unique structural features of CheR, such as the beta subdomain, are probably necessary for CheR's specific interaction with its substrates, the bacterial chemotaxis receptors.

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Escherichia coli aspartate receptor. Oligomerization of the cytoplasmic fragment

Cited by 9 publications

References 11 publications

Demonstration of dimer formation of the cytoplasmic domain of a transmembrane osmosensor protein, EnvZ, of Escherichia coli using Ni‐histidine tag affinity chromatography

Demonstration of dimer formation of the cytoplasmic domain of a transmembrane osmosensor protein, EnvZ, of Escherichia coli using Ni‐histidine tag affinity chromatography

Membrane Association of a Protein Increases the Rate, Extent, and Specificity of Chemical Cross-Linking

Crystal structure of the chemotaxis receptor methyltransferase CheR suggests a conserved structural motif for binding S-adenosylmethionine

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