Quantitative Microdetermination of Enzymes in Sweat Gland III. Succinic Dehydrogenase in Cystic Fibrosis.

Gibbs, Gordon E.; Reimer, Keith A.

doi:10.3181/00379727-119-30213

Cited by 26 publications

(9 citation statements)

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“…The plasma membranes were assayed by previously described methods for the marker enzymes Na,K-ATPase, a basal-lateral membrane marker (Schoner, von Ilberg, Kramer & Seubert, 1967), alkaline phosphatase, a proximal tubule luminal membrane enzyme (Bessey, Lowry & Brock, 1947) and perhaps also found in the luminal membranes of the TALH (Bonting, Pollack, Muehrcke & Kark, 1958), filipin-sensitive Mg ++-ATPase (Kinne-Saffran & Kinne, 1979), a potential luminal enzyme marker for the TALH, NADH dehydrogenase (Wallach & Kamat, 1966), an endoplasmic reticuhim enzyme and succinic dehydrogenase (Gibbs & Reimer, 1965), a mitochondrial enzyme marker. Protein was measured by the method of Lowry, Rosebrough, Farr and Randall (1951) after precipitation by 10% trichloracetic acid (wt/vol).…”

Section: Preparation Of the Plasma Membrane Fractionmentioning

confidence: 99%

Sodium-chloride transport in the medullary thick ascending limb of henle's loop: Evidence for a sodium-chloride cotransport system in plasma membrane vesicles

Eveloff

Kinne

1983

J. Membrain Biol.

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Sodium transport mechanisms were investigated in plasma membrane vesicles prepared from the medullary thick ascending limb of Henle's loop (TALH) of rabbit kidney. The uptake of 22Na into the plasma membrane vesicles was investigated by a rapid filtration technique. Sodium uptake was greatest in the presence of chloride; it was reduced when chloride was replaced by nitrate, gluconate or sulfate. The stimulation of sodium uptake by chloride was seen in the presence of a chloride gradient directed into the vesicle and when the vesicles were equilibrated with NaCl, KCl plus valinomycin so that no chemical or electrical gradients existed across the vesicle (tracer exchange experiments). Furosemide decreased sodium uptake into the vesicles in a dose-dependent manner only in the presence of chloride, with a Ki of around 5 X 10(-6) M. Amiloride, at 2 mM, had no effect on the chloride-dependent sodium uptake. Similarly, potassium removal had no effect on the chloride-dependent sodium uptake and furosemide was an effective inhibitor of sodium uptake in a potassium-free medium. The results show the presence of a furosemide-sensitive sodium-chloride cotransport system in the plasma membranes of the medullary TALH. There is no evidence for a Na+/H+ exchange mechanism or a Na+ -K+ -Cl- cotransport system. The sodium-chloride cotransport system would effect the uphill transport of chloride against its electrochemical potential gradient at the luminal membrane of the cell.

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Section: Preparation Of the Plasma Membrane Fractionmentioning

confidence: 99%

Sodium-chloride transport in the medullary thick ascending limb of henle's loop: Evidence for a sodium-chloride cotransport system in plasma membrane vesicles

Eveloff

Kinne

1983

J. Membrain Biol.

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“…The assay for succinic dehydrogenase activity was performed after preincubation in 0.1% desoxycholate for 30 rain at 0 ~ according to the method of Gibbs and Reimer [19]. Cytochrorne c oxidase was determined according to the method of Cooperstein and Lazarow [8].…”

Section: Enzymatic Characterization Of the Membrane Fraction And Protmentioning

confidence: 99%

An ATP-driven proton pump in brush-border membranes from rat renal cortex

1982

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The rate of ATP hydrolysis in ATP-preloaded plasma membrane vesicles derived from the luminal membrane of renal cortical tubules, and the rate of H+ secretion out of the same vesicles were investigated. Both were inhibited at low temperature, by the action of filipin, an antibiotic that complexes with cholesterol in plasma membranes, and by the action of blockers of mitochondrial Fo hydrogen channels, dicyclohexylcarbodiimide and Dio-9. Valinomycin in the presence of K+ showed a stimulatory effect, the protonophor carbonyl-cyanid-p-trifluormethoxy-phenylhydrazone stimulated the intravesicular ATP hydrolysis and apparently abolished acidification of the extravesicular medium. Lowering of the pH of the extravesicular medium retarded ATP hydrolysis, while readjustment of extra- and intravesicular pH accelerated ATP hydrolysis again. These findings strongly support the assumption that an ATP-driven proton pump is located in the luminal membrane of renal cortical tubules.

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“…The values were corrected for the substrate blank and for the activity found in the absence of any divalent cation. The succinic dehydrogenase was measured according to Gibbs and Reimer [9] after pretreatment of the samples with 0.1% desoxycholate.…”

Section: Protein and Enzyme Assaymentioning

confidence: 99%

Localization of a calcium-stimulated ATPase in the basal-lateral plasma membranes of the proximal tubule of rat kidney cortex

1974

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Summary. In the proximal tubule of the kidney, calcium is reabsorbed by an active transport mechanism. Recently, a Ca2+-activated ATP-phosphohydrolase has been described in plasma membranes from rat kidney cortex, which is different from the CaZ+-ATPase present in mitochondria. To elucidate the role of this enzyme in transepithelial calcium transport we studied its localization within the cell of the proximal tubule. For this purpose, a plasma membrane fraction was subdivided by preparative free flow electrophoresis and the distribution of alkaline phosphatase (brush border microvillus membranes) and Na + -K+-ATPase (basal-lateral plasma membranes) was compared with that of Ca2+-ATPase. The distribution pattern obtained and the corresponding enrichment factors show that a nonmitochondrial Ca2+-ATPase is--in analogy to the Na + --K+-ATPase--located only in the basal-lateral plasma membranes of the proximal tubule. Regarding the different substrate specificity and the insensitivity of the enzyme towards sodium, potassium and ouabain it seems to be possible to differentiate between the two enzymes at a molecular level. It is proposed that the Ca a+-stimulated ATPase is involved in the active transtubular transport of calcium.In the proximal tubule of the kidney, besides monovalent cations like sodium and potassium, the bivalent cation calcium is also reabsorbed from the primary urine [28]. The transport of calcium is performed against a concentration gradient and has many similarities with the sodium transport [8]. Thus, the kidney plays an important role in the regulation of the homeostasis of the divalent cation [26] which is essentially involved in processes like neuronal transmission, permeability of cell membranes, cell adhesion and bone formation [5].The biochemical events taking place when calcium is transported through the cell have been studied extensively in the gut, mainly by the group of * A preliminary report of this work was presented at the Fall Meeting of the Deutsche Physiologische Gesellschaft, Dtisseldorf 1972.

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Quantitative Microdetermination of Enzymes in Sweat Gland III. Succinic Dehydrogenase in Cystic Fibrosis.

Cited by 26 publications

References 0 publications

Sodium-chloride transport in the medullary thick ascending limb of henle's loop: Evidence for a sodium-chloride cotransport system in plasma membrane vesicles

Sodium-chloride transport in the medullary thick ascending limb of henle's loop: Evidence for a sodium-chloride cotransport system in plasma membrane vesicles

An ATP-driven proton pump in brush-border membranes from rat renal cortex

Localization of a calcium-stimulated ATPase in the basal-lateral plasma membranes of the proximal tubule of rat kidney cortex

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