Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.6.1.3 (ATPase)
 65,361 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

In dog proximal tubules in suspension, the addition of glucose increased significantly the ouabain-sensitive fraction of respiration, a response suppressed by phlorizin. The addition of alpha-methyl-D-glucoside (alpha-MG) had a modest effect and 3-O-methyl-D-glucoside (3-O-MG) had no effect. The different stimulation of the Na+,K(+)-ATPase activity elicited for each hexose could be explained by a different increment of net transepithelial flux of sodium induced by the sodium: hexose cotransport. This flux is a direct function of the transport characteristics of both luminal and antiluminal membranes of proximal cells for these sugars: glucose is rapidly transported by both membranes (allowing a large transepithelial flux of glucose: sodium) while alpha-MG is poorly transported by the basolateral, and 3-O-MG by the luminal, membrane of the dog proximal tubule (allowing a small transepithelial flux of hexoses and sodium). However the overall tubular respiration of dog proximal tubules was not increased by glucose addition because the increment in the ouabain-sensitive fraction was accompanied by a reciprocal decrement in an ouabain-insensitive but oligomycin- or N',N' dicyclohexylcarbodiimide (DCCD)-sensitive (or in the bafilomycin-sensitive) component of respiration. This component reflects the activity of a large BBM-bound H(+)-ATPase found in this species. The intracellular pH of dog proximal tubules in suspension was measured using the proton-sensitive fluorescent probe 2',7'-bis-2-(carboxyethyl)-5, (and 6)-carboxyfluorescein. Glucose application significantly alkalinized the cells. In contrast, other substrates such as lactate or acetate simultaneously acidified the cells and increased the ouabain-insensitive phosphorylative respiration of dog tubules. These observations suggest that a modulation of the activities of both the sodium and most probably the proton pump is elicited by substrate availability in suspensions of proximal tubules.
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PMID:Substrate-induced modulation of ATP turnover in dog and rabbit proximal tubules. 132 87

The antiulcerogenic drug ranitidine, given orally to mice, brought about reductions of kidney-bound hydrolytic enzymes at three different dose levels, viz. 10 mg, 100 mg, and 1000 mg/kg body weight, and for three different time points (single administration for 2 h and 24 h, and daily administration for 15 days). The activities of Na+, K(+)-ATPase, Ca2(+)-ATPase, and Mg2(+)-ATPase (marker enzymes of basolateral membranes) were reduced, and these reductions were significant at higher doses and after a 24-h single treatment or 15 days' daily treatment. Maltase, alkaline phosphatase, and leucine aminopeptidase (marker enzymes of brush border membrane [BBM]) activities were significantly inhibited after ranitidine treatment. Kinetic analysis of BBM-associated enzymes indicated that ranitidine decreased the maximum of apparent initial enzyme velocity (Vmax) of maltase, alkaline phosphatase, and leucine aminopeptidase. The substrate affinity constant (Km) was decreased in the case of alkaline phosphatase and maltase, while it was not altered in the case of leucine aminopeptidase. In vitro addition of ranitidine to renal BBM also produced significant inhibition of these enzymes, the inhibition constants (Ki) for maltase, alkaline phosphatase, and leucine aminopeptidase being 7.5, 15.5, and 3.5 mM, respectively. Membrane-bound lipid estimation showed a significant increase in phospholipids, triglycerides, and free fatty acids. Cholesterol, however, was decreased in both renal basolateral and brush border membranes.
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PMID:Effect of histamine H2-receptor antagonist, ranitidine on renal brush border and basolateral membranes. 217 15

To determine if ischemia induces alterations in renal proximal tubule surface membranes, brush border (BBM) and basolateral membranes (BLM) were isolated simultaneously from the same cortical homogenate after 50 min of renal pedicle clamping. Ischemia caused a selective decrease in the specific activity of BBM marker enzymes leucine aminopeptidase and alkaline phosphatase, but did not effect enrichment (15 times). Neither specific activity nor enrichment (10 times) of BLM NaK-ATPase was altered by ischemia. Contamination of BBM by intracellular organelles was also unchanged, but there was an increase in the specific activity (41.1 vs. 60.0, P less than 0.01) and enrichment (2.3 vs. 4.3, P less than 0.01) of NaK-ATPase in the ischemic BBM fraction. Ischemia increased BLM lysophosphatidylcholine (1.3 vs. 2.5%, P less than 0.05) and phosphatidic acid (0.4 vs. 1.3%, P less than 0.05). Ischemia also decreased BBM sphingomyelin (38.5 vs. 29.6%, P less than 0.01) and phosphatidylserine (16.1 vs. 11.4%, P less than 0.01), and increased phosphatidylcholine (17.2 vs. 29.7%, P less than 0.01), phosphatidylinositol (1.8 vs. 4.6%, P less than 0.01), and lysophosphatidylcholine (1.0 vs. 1.8%, P less than 0.05). The large changes in BBM phospholipids did not result from new phospholipid synthesis, since the specific activity (32P dpm/nmol Pi) of prelabeled individual and total phospholipids was unaltered by ischemia. We next evaluated if these changes were due to inability of ischemic cells to maintain surface membrane polarity. Cytochemical evaluation showed that while NaK-ATPase could be detected only in control BLM, specific deposits of reaction product were present in the BBM of ischemic kidneys. Furthermore, using continuous sucrose gradients, the enzymatic profile of ischemic BBM NaK-ATPase shifted away from ischemic BLM NaK-ATPase and toward the BBM enzymatic marker leucine aminopeptidase. Taken together, these data suggest that NaK-ATPase activity determined enzymatically and cytochemically was located within ischemic BBM. We propose that ischemia impairs the ability of cells to maintain surface membrane polarity, and also results in the accumulation of putative calcium ionophores.
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PMID:Ischemia induces partial loss of surface membrane polarity and accumulation of putative calcium ionophores. 300 Nov 41

Renal proximal tubule cells adapt to dietary phosphate (Pi) restriction by increasing Pi transport independent of parathyroid hormone, vitamin D metabolites, or serum Ca2+. To determine the underlying cellular mechanism(s), brush border (BBM) and basolateral membranes (BLM) were isolated from growing male rats fed a synthetic diet containing variable levels of Pi (0.1-1.4%). Dietary Pi restriction was without effect on either BBM or BLM total lipid phosphorus, individual phospholipid species, or BLM Na+-K+-ATPase specific activity. However, dietary Pi restriction (0.1 vs. 1.0%) did cause a significant reduction in BBM but not BLM cholesterol (0.45 vs. 0.41 mumol/mg protein). Brush border membrane cholesterol was inversely correlated with the tubular reabsorption of Pi (r = 0.77, P less than 0.01) over a broad range (99.9-46.2%). Arrhenius analysis of two intrinsic BBM enzymes revealed a significant reduction in the breakpoint temperature for alkaline phosphatase but no change for Mg2+-ATPase. Fluorescence polarization studies showed increased BBM inner core fluidity due to an alteration in neutral lipids but not phospholipid, fatty acid, or protein membrane components. These data demonstrate that the BBM can regulate its cholesterol content independent of the BLM. Furthermore, they suggest that adaptation to dietary Pi restriction involves a reduction in BBM cholesterol, which may be mediated by an increase in membrane fluidity.
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PMID:Renal apical membrane cholesterol and fluidity in regulation of phosphate transport. 316 Feb 47

Previous studies have implicated glucocorticoids as an important factor in the postnatal maturational increase in proximal tubule volume absorption, Na+/H+ antiporter, Na(HCO3)3 symporter, and Na(+)-K(+)-ATPase activity. The present study examined whether glucocorticoids are also a potentially important factor in the maturational decrease in proximal tubule phosphate transport. Renal BBMs were prepared from neonatal rabbits who received dexamethasone (10 micrograms/100 g body weight) or vehicle. Brush-border membrane vesicles from dexamethasone-treated neonates had a lower rate of Na-phosphate cotransport than controls (50.8 +/- 3.6 versus 29.2 +/- 2.6 pmol 32P(i)/10 s/mg protein, p < 0.001). This decrease was due to a decrease in the Vmax with no change in the affinity of the transporter for phosphate. The dexamethasone-induced decrease in BBM Na-phosphate transport was not due to a reduction in transporters as assayed by phosphate-protectable Na-dependent equilibrium binding of phosphonoformic acid. Dexamethasone treatment caused an increase in the fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene and trimethylammonium-1,6-diphenyl-1,3,5-hexatriene (i.e. a decrease in membrane fluidity). Brush-border membranes from dexamethasone-treated neonates had a decrease in sphingomyelin and an increase in phosphatidylcholine and phosphatidylinositol content but no change in cholesterol or total phospholipid content. These data are consistent with glucocorticoids playing a role in the postnatal maturational decrease in proximal tubule phosphate transport by altering membrane characteristics.
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PMID:Maturational effects of glucocorticoids on neonatal brush-border membrane phosphate transport. 804 84

Brush border (BBM) and basolateral membranes (BLM) of rat renal cortical cells separated by free flow electrophoresis revealed two distinct peaks of BBM-specific leucine aminopeptidase and Na+/K(+)-ATPase for BLM. PTH/PTH-related protein (PTHrP) receptors were identified in BBM and BLM. Specific binding of 125 pM [125I]chicken [Tyr36]-PTHrP-(1-36)amide [chPTHrP-(1-36)] to individual fractions of membranes separated by free flow electrophoresis overlapped with the leucine aminopeptidase and Na+/K(+)-ATPase profiles. Binding to pooled BBM was 53 +/- 5% (mean +/- SEM) of that to BLM (P < 0.01). In BBM and BLM, half-maximal inhibition of binding was obtained with 0.4-0.9 nM chPTHrP-(1-36) and 0.2-0.6 nM rat PTH-(1-34). Guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S; 100 microM) lowered chPTHrP-(1-36) binding to 50% of control levels, and half-maximal inhibition of binding was obtained with 480 and 8 nM GTP gamma S in BBM and BLM, respectively. Cross-linking of the PTH/PTHrP receptors with [125I]chPTHrP-(1-36) modified with N-hydroxysuccinimidyl-4-azidobenzoate revealed indistinguishable doublets of 83 and 73 kilodaltons in both BBM and BLM. Adenylyl cyclase was stimulated 6- and 10-fold by chPTHrP-(1-36) and GTP gamma S, respectively, in BLM and 1.3- and 1.9-fold in BBM. In conclusion, PTH receptors were recognized in both the basolateral and brush border membranes. Different receptor coupling to G-proteins and minimal cAMP stimulation in BBM provide evidence for PTH/PTHrP receptor isotypes and/or different postreceptor activation in BBM and BLM.
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PMID:Apical and basolateral parathyroid hormone receptors in rat renal cortical membranes. 811 56

To examine the contribution of exogenous calcium and oxalate in magnesium deficiency, intestinal absorption of both calcium and oxalate was studied by preparing brush border membrane vesicles (BBMV). Purity of the BBMV was ascertained biochemically by enrichment of the marker enzyme alkaline phosphatase by 14-fold with a concomitant 90 per cent decrease in the basolateral marker enzyme Na+/K(+)-ATPase in the purified membrane preparation as compared to the respective homogenate in both the groups. Uptake studies were carried out by a rapid filtration technique. The kinetics were studied by measuring the rate of influx as a function of concentration (0.1-1.0 mM). BBMV from both the groups showed a linear positive relationship between the uptake rate and the concentration for both calcium and oxalate, thereby demonstrating that calcium and oxalate are transported through intestinal microvillus membrane by a simple passive diffusion process. However, the rate of uptake of calcium and oxalate was significantly higher in the magnesium-deficient group as compared to the pair-fed control group, as shown by the increase in the slope line for both calcium and oxalate (for calcium, control = 3.88, deficient = 5.86; for oxalate, control = 4.41, deficient = 7.20). Analysis of the lipid composition of the BBM revealed a significant decrease in the cholesterol content (P < 0.01) with a concomitant increase in the triglycerides (P < 0.01) and total fatty acid content (P < 0.001) in the magnesium-deficient group. Thus the results indicate that, although the mechanism of translocation of calcium and oxalate in the intestine is similar in the two groups, magnesium deficiency leads to hyperabsorption of both the ligands through alterations in the lipid composition of the membrane, thereby increasing the risk of stone formation.
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PMID:Intestinal absorption of calcium and oxalate in magnesium-deficient rats. 839 6

During the preparation of a suspension of dog kidney proximal tubules by collagenase treatment, an uptake of FITC-albumin was demonstrated. This process is attributed to the activation of receptor-mediated endocytosis leading to the appearance of FITC-albumin into intracellular vesicular structures. The isolation of brush border membrane vesicles (BBMV) from the dog kidney proximal tubules in suspension by the magnesium precipitation technique leads to the copurification of a large population of endosomes. These endosomes were separated from BBM vesicles by a technique involving wheat-germ agglutinin. The enrichment in BBM markers and in bafilomycin-sensitive ATPase activity was comparable in endosomes and BBM vesicles. However, the acridine orange acidification assay showed a V-type ATPase-dependent acidification in endosomes but not in BBMV, demonstrating a different orientation of the proton pumps in these structures. SDS-PAGE analysis also showed significant differences in protein pattern of vesicles and endosomes. The most notable difference was the presence of 42-44 kDa and 20-24 kDa proteins in BBMV and their complete absence in endosomes. Western blot analysis identified these proteins as actin and RhoA, among other small proteins, respectively. Western blot experiments also demonstrated a different distribution of beta-COP, beta-adaptin, and RhoGDI in vesicles and endosomes. The morphological aspect (electron microscopy) and sedimentation of endosomes in a 50% Percoll gradient identified these structures as "heavy endosomes" (buoyant density D = 1.036 g/ml). Flow cytometry analysis of heavy endosomes purified from tubules isolated in presence of FITC-albumin showed the presence of FITC-albumin in up to 92% of these intracellular organelles. Western blot analysis using anti-FITC and anti-collagenase antibodies allowed quantification of the FITC-albumin and collagenase A in the purified endosomes. Our results indicate that heavy endosomes are formed during the preparation of the proximal tubules following activation of receptor-mediated endocytosis, probably by soluble proteins. The suspension of tubules thus offers a experimental tool to study the protein reabsorption and traffic of endosomal vesicles in the proximal tubules.
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PMID:Isolation of heavy endosomes from dog proximal tubules in suspension. 869 8

Active glycine transport was demonstrated in microvillous (maternal-facing, BBM) and basal (fetal-facing, BCM) plasma membranes of the human term placental syncytiotrophoblast. The kinetic studies showed that the amino acid had a distinct overshoot at 1 min in BBM and 3 min in BCM vesicles while a steady state rate was achieved in approx 5 min in both the vesicles. Glycine transport is highly ion-specific and its dependency on Na+ can not be satisfied by replacing with other monovalent cation. Cl- is also implicated in the generation of the electrochemical gradient and replacement of Cl- with SO4(2-) anions failed to stimulate the transport process. The transport process was saturable with external glycine which exhibited rectangular hyperbolic kinetics typical of a mediated movement. The calculated kt and Jmax from the linear transformation of the data were 6.67 & 4 mM and 294 & 263 nmoles glycine. mg protein-1.min-1 in the BBM and BCM vesicles, respectively. The glycine transport was inhibited by a number of other amino acids which are known to be transported through the A and ASC systems. The glycine transport system may be dependent on multiple pathways such as the A, ASC or Gly which is a variant of pathway A. Glycine transport was inhibited by ouabain, a known Na+/K+ -ATPase inhibitor, in the BCM vesicles but not in the BBM system. Nicotine, insulin, sodium fluoride and sodium arsenate were inhibitors for both the vesicles.
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PMID:Transport of glycine in the brush border and basal cell membrane vesicles of the human term placenta. 893 15

The purpose of the present study is to analyze membrane fluidity, enzyme, phospholipid and fatty acid composition and cholesterol content in the brush border (BBM) and basolateral (BLM) membranes obtained from the renal cortex of normal dogs. All measurements were carried out in samples from the same kidney in order to correlate membrane fluidity with membrane composition. BBM and BLM were obtained separately by MgCl precipitation and gradient centrifugation. The order parameter of membrane fluidity was measured by 1,6-dimethyl-1,3,5-hexatriene (DPH) and 1-trimethylammoniophenyl-DPH. (TMA-DPH) steady-state polarization fluorescence. Total lipids, phospholipids and phospholipid classes, cholesterol content, and fatty acid classes were also measured. Data from BLM enzymatic activity revealed an 11-fold enrichment in Na,K-ATPase, whereas the enrichment factors for the other enzymatic markers were well below the unit, demonstrating the high purity of the preparation obtained. Similarly, BBM showed a 9 times increase in alkaline phosphatase and gamma-glutamyltranspeptidase enrichment, and values of enrichment factors for the other enzymatic markers of about 1. BBM exhibited a higher value of steady-state fluorescence anisotropy and thus a lower fluidity than BLM using either of the fluorescent probes DPH or TMA-DPH. This lower fluidity in both the central hydrophobic zone, and the fluorescent probes DPH or TMA-DPH. This lower fluidity in both the central hydrophobic zone, and the external, more hydrophilic leaflet of BBM in comparison with BLM was consistent with the findings of: (a) a higher cholesterol/protein ratio; (b) a lower phospholipid protein ratio; (c) a higher sphingomyelin/choline glycerophospholipid ratio, and (d) a lower unsaturation degree of the fatty acids.
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PMID:Biochemical and functional characterization of renal cortical brush border and basolateral membranes in dogs. 895 34


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