The KDP ATPase of Escherichia coli.
Explore the source record for details and available documents.
Adenosine Triphosphatases↗
Biomedical subjects
W Epstein
Publications and source records attributed to W Epstein.
Purification and kinetic characterization of the Kdp-ATPase.
Explore the source record for details and available documents.
Adenosine Diphosphate↗
Attentional demands of processing shape in three-dimensional space: evidence from visual search and precuing paradigms.
The hypothesis that representation of projective shape is preattentive whereas representation of objective shape in three-dimensional space requires allocation of attention was tested in 2 visual search and 2 precuing experiments. In the visual search experiments, the slope for projective shape search was expected to approach 0 and that for objective shape search was expected to be a positive monotonic function of set size. In the precuing experiments, the effects of precuing were expected to be largely limited to the task requiring representation of objective shape. The overall pattern of results conformed to expectations. The findings are interpreted in the context of a model of shape-at-a-slant processing set out by Epstein and Lovitts (1985) and Epstein and Babler (1989, 1990).
Adult↗
The products of the kdpDE operon are required for expression of the Kdp ATPase of Escherichia coli.
The expression of the Kdp system for K+ uptake in Escherichia coli requires the products of two genes, kdpD and kdpE. These genes constitute an operon adjacent to the kdpABC operon that encodes the three membrane protein subunits of Kdp. Both operons are transcribed in the same direction and overlap; the kdpDE promoter is in kdpC, the last gene of the kdpABC operon. Transcription of the kdpDE operon is at a low level when Kdp is not expressed; transcription increases about 10-fold when kdpABC is turned on, indicating significant read-through of the kdpDE operon by transcripts beginning at the promoter of kdpABC operon. The proximal region of the kdpD gene is the site of most mutations that lead to constitutive expression of the kdpABC operon.
Adenosine Triphosphatases↗
KdpD and KdpE, proteins that control expression of the kdpABC operon, are members of the two-component sensor-effector class of regulators.
The Kdp system of Escherichia coli, a transport ATPase with high affinity for potassium, is expressed when turgor pressure is low. Expression requires KdpD, a 99-kDa membrane protein, and KdpE, a 25-kDa soluble cytoplasmic protein. The sequences of KdpD and KdpE show they are members of the sensor-effector class of regulatory proteins: the C-terminal half of KdpD is homologous to sensors such as EnvZ and PhoR, and KdpE is homologous to effectors such as OmpR and PhoB. The predicted structure of KdpD suggests that it is anchored to the membrane by four membrane-spanning segments near its middle, with both C- and N-terminal portions in the cytoplasm. Subcellular fractionation confirms the expected location of the protein in the inner membrane. The N-terminal region has no homology to known proteins and is the site of mutations that make Kdp expression partially constitutive; this portion may serve to sense turgor pressure. Since several other sensor-effectors have been shown to mediate control through phosphorylation, this mechanism is proposed to control expression of Kdp.
Adenosine Triphosphatases↗
Kdp, a bacterial P-type ATPase whose expression and activity are regulated by turgor pressure.
The Kdp ATPase is a P-type ATPase consisting of three large protein subunits in a complex that probably contains 2 copies of each subunit. A small hydrophobic peptide, encoded in the same operon as the large subunits, may also participate. Kdp has very high affinity for K+ and serves to scavenge this ion when its concentration is low. Kdp responds to turgor pressure at two levels, at the level of activity and of expression. Kdp mediates net uptake when turgor is low, but mediates exchange without net change when turgor is normal. Kdp is expressed only when turgor is low. This control is mediated by a pair of regulatory proteins, members of the class of 'sensor-effector' regulators widely distributed in bacteria. It is suggested that low turgor changes the conformation of the KdpD 'sensor' protein, activating its presumed kinase activity to phosphorylate KdpE, the 'effector' protein, and phospho-KdpE in turn turns on expression of the operon that encodes the Kdp complex.
Adenosine Triphosphatases↗
Treatment of oral hairy leukoplakia with podophyllin.
Explore the source record for details and available documents.
Administration, Topical↗
Escherichia coli accumulates the eukaryotic osmolyte taurine at high osmolarity.
Escherichia coli accumulated taurine at high osmolarity via the ProU and ProP transport systems. Taurine accumulation was shown to be osmotically active as it displaced cytoplasmic K+. In contrast to betaine and proline, taurine only modestly enhanced the growth rate of E. coli at high osmolarity and only if the cell was unable to synthesise trehalose. These studies show that taurine cannot be used as a tracer for the extra-cytoplasmic space of bacteria grown at high osmolarity.
Amino Acid Transport Systems↗
Genetic analysis of potassium transport loci in Escherichia coli: evidence for three constitutive systems mediating uptake potassium.
The analysis of mutants of Escherichia coli that require elevated concentrations of K+ for growth has revealed two new genes, trkG, near minute 30 within the cryptic rac prophage, and trkH, near minute 87, the products of which affect constitutive K+ transport. The analysis of these and other trk mutations suggests that high rates of transport, previously considered to represent the activity of a single system, named TrkA, appear to be the sum of two systems, here named TrkG and TrkH. Each of these two is absolutely dependent on the product of the trkA gene, a cytoplasmic protein associated with the inner membrane (D. Bossemeyer, A. Borchard, D. C. Dosch, G. C. Helmer, W. Epstein, I. R. Booth, and E. P. Bakker, J. Biol. Chem. 264:16403-16410, 1989). The TrkH system is also dependent on the products of the trkH and trkE genes, while the TrkG system is also dependent on the product of the trkG gene and partially dependent on the product of the trkE gene. It is suggested that the trkH and trkG products are membrane proteins that form the transmembrane path for the K+ movement of the respective systems. Two mutations altering the trkA product reduce the affinity for K+ of both TrkG and TrkH, indicating that changes in peripheral protein can alter the conformation of the sites at which K+ is bound prior to transport. The TrkD system has a relatively modest rate of transport, is dependent solely on the product of the trkD gene, and is the sole saturable system for Cs+ uptake in this species (D. Bossemeyer, A. Schlösser, and E. P. Bakker, J. Bacteriol. 171:2219-2221, 1989).
Biological Transport, Active↗
The bacterial Kdp K(+)-ATPase and its relation to other transport ATPases, such as the Na+/K(+)- and Ca2(+)-ATPases in higher organisms.
The Kdp system is a three-subunit member of the E1-E2 family of transport ATPases. There is sequence homology of the 72 kDa KdpB protein, the largest subunit of Kdp, with the other members of this family. The predicted structure of the 21 kDa KdpC subunit resembles that of the beta subunit of the Na+,K(+)-ATPase, suggesting that these subunits may have a similar function. The 59 kDa KdpA subunit has no known homologue; it is very hydrophobic and is predicted to cross the membrane 10-12 times. Genetic studies implicate this subunit in the binding of K+. As the binding site must be close to the beginning of the transmembrane channel, we suggest that KdpA also forms most or all of the latter. KdpA may have evolved from a K+/H+ antiporter that was recruited by the KdpB precursor to achieve the high affinity and specificity for K+, and the activation of transport by low turgor pressure characteristic of Kdp. Turgor pressure controls the expression of Kdp. This action is dependent on the 70 kDa KdpD and 23 kDa KdpE proteins. We are in the process of sequencing these genes. KdpE is homologous to the smaller protein of other members of a family of pairs of regulatory proteins implicated in control of a variety of bacterial processes such as porin synthesis, phosphate regulon expression, nitrogen metabolism, chemotaxis and nodule formation.
Adenosine Triphosphatases↗
Suppression of urushiol-induced delayed-type hypersensitivity responses in mice with serum IgG immunoglobulin from human hyposensitized donors.
Serum IgG immunoglobulin fractions from human subjects hyposensitized to poison ivy/oak by oral administration of urushiol suppressed the induction of delayed-type hypersensitivity (DTH) responses in mice to this hapten. This suppressive activity was hapten specific because it did not modify DTH responses to dinitrofluorobenzene (DNFB). Absorption of human serum with lymph node cells from urushiolsensitized but not DNFB-sensitized mice removed the suppressive activity, suggesting that anti-idiotypic antibodies reacting with T-cell receptors are involved.
Animals↗
The gdhA gene is located at 38.6 minutes on the Escherichia coli map.
We report here that the gdhA gene of Escherichia coli, which encodes the NADP-specific glutamate dehydrogenase, is located at 38.6 min on the map. We have confirmed this location by showing linkage with three Tn10 insertions that are linked to the aroD, pheS, and ansA loci, by complementation by a restriction-mapped lambda clone, and by showing correspondence between the restriction maps of the chromosome and the cloned and sequenced gdhA gene.
Chromosomes, Bacterial↗
Involvement of gamma-glutamyl peptides in osmoadaptation of Escherichia coli.
Accumulation of K+ ions and glutamate plays a primary role in maintaining osmotic balance in Escherichia coli, as illustrated by the high concentrations of these ions present in cells growing in medium of high osmolality. We found that two gamma-glutamyl peptides and glutamine also accumulated during growth at high osmolarity. In a mutant unable to make trehalose growing in 1.3 osM medium, glutathione, gamma-glutamylglutamine, and glutamine accumulated to levels of 73, 33, and 140 mumol/g of protein, respectively. In such cells, K+ was present at 1,450 mumol/g of protein, indicating that glutathione and gamma-glutamylglutamine accounted for less than 10% of the low-molecular-weight anions accumulated with K+. However, glutathione is needed for wild-type osmotolerance in this species. A mutant deficient in glutathione because of an insertion in the gshA gene was unable to grow above 1.4 osM, grew more slowly at intermediate osmolarities, and took longer to adapt to growth following osmotic upshock. The involvement of glutathione in osmoregulation was independent of the effect of glutathione on K+ retention.
Chromatography, High Pressure Liquid↗
In search of depth.
Four experiments designed to investigate a visual search task are reported. In each experiment, subjects searched for either a two-dimensional shape rotated in depth among frontal-parallel distractors or a frontal-parallel shape among distractors that were rotated in depth. The principal independent variable was search-set size. In addition, over the four experiments, a variety of spatial arrangements and two axes of rotation in depth were sampled. The chief aim of the experiments was to adduce evidence bearing on the attentional demands of searching for depth. The slopes of the reaction-time (RT) functions were taken as diagnostic. Experiments 1-3 exhibited positive slopes for the RT-set-size function. These slopes appear to be due to a conscious adoption of a serial search strategy by the subjects. When this tendency was suppressed by the procedures of Experiment 4, the slope of the RT-set-size functions did not differ significantly from zero. We conclude that, in agreement with the findings of other studies, slant-in-depth can be detected preattentively.
Adult↗
An asymmetry in transmodal perceptual learning.
Transmodal perceptual learning was examined in a canonical same/different paradigm. Subjects naive to vibrotactile stimulation and unfamiliar with the task were asked to discriminate sequentially presented shapes moving laterally across an aperture. On all trials, the shapes were presented either visually and then vibrotactually or in the opposite modality order, and on all trials the shapes moved in opposite directions. Analyses of the data revealed that although perceptual learning was evident in both groups, the rate of the learning was more rapid in the visual-vibrotactile group. This interaction of modality order and practice was significant and was considered in terms of E.J. Gibson's theory of perceptual learning and in terms of the constructs, suggested by J.J. Gibson, of available and accessible information-in-stimulation and the education of attention.
Adult↗
K+-transport protein TrkA of Escherichia coli is a peripheral membrane protein that requires other trk gene products for attachment to the cytoplasmic membrane.
The TrkA protein, which is essential for the activity of the constitutive Trk K+-uptake system of Escherichia coli, is a peripheral membrane protein. The protein was detected in immunoblots by polyclonal antibodies to sodium dodecyl sulfate-denatured TrkA protein. In extracts from wild-type cells equal amounts of TrkA were found in the membrane and soluble fractions, suggesting that membrane binding is relatively weak. When the protein was moderately overproduced it appeared mainly in the soluble fraction; stronger overproduction led to the formation of aggregates that could not be solubilized by nonionic detergents. Mutations in the three other genes implicated in Trk activity, trkE, trkG, and trkH, reduced or abolished the binding of TrkA to the membrane. These results support the model, previously based solely on genetic data, that Trk is a multisubunit complex and implicates the products of the other trk genes in the normal binding of TrkA to the complex in the cytoplasmic membrane.
Antibodies↗
Perceptual learning of spatiotemporal events: evidence from an unfamiliar modality.
Perceptual learning was examined in two experiments in which subjects, originally unfamiliar with vibrotactile stimulation, were required to identify dynamic vibrotactile patterns with static visual patterns of the same two-dimensional shapes. In Experiment 1 we examined changes in performance with practice under a variety of vibrotactile spatial and temporal conditions. In Experiment 2 we investigated transfer of learning from one set of vibrotactile patterns to another different set. In neither experiment were subjects supplied with knowledge of results. Substantial perceptual learning (improvement in identification with practice) was observed in Experiment 1, although a minority of subjects did not exhibit improvement. Experiment 2 confirmed the general findings of Experiment 1 and also provided evidence of substantial positive transfer. In both experiments, multidimensional scaling of pattern confusion data revealed that practice (and improvement in identification) did not qualitatively change the relative confusability of patterns, suggesting that the (informative) structure of the patterns, irrespective of familiarity with a specific set of patterns, determined confusability. The findings are interpreted in terms of learning constructs offered by E. J. and J. J. Gibson. We conclude by considering the prospects that a connectionist mechanism can account for the observed perceptual learning.
Attention↗
Wide distribution of homologs of Escherichia coli Kdp K+-ATPase among gram-negative bacteria.
We used Southern blotting to screen a variety of bacterial genes for homology to the kdp genes of Escherichia coli, genes that encode an ATP-driven K+ transport system. We found that most enterobacteria have sequences homologous to those of the three kdp structural genes and the kdpD regulatory gene. A number of distantly related species, including some cyanobacteria, have sequences homologous to those of the structural genes but not the regulatory gene. In all cases only a single region of homology was found. These results suggest that ATP-driven transport systems similar to the Kdp system in structure and regulation are found in many enteric organisms. In other gram-negative organisms, the ATPase is more divergent, retaining good homology at the DNA level only to the highly conserved phosphorylated subunit of the ATPase.
Adenosine Triphosphatases↗