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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
1
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pubmed:dateCreated |
1990-5-16
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pubmed:abstractText |
Thiazide-type diuretics act at receptors to inhibit NaCl transport in the renal distal tubule. We solubilized high-affinity [3H]metolazone binding sites from rat kidney membranes with Triton X-100, which was more effective than several other detergents. Phosphatidylcholine and a mixture of proteinase inhibitors were needed to stabilize binding so that 57% of solubilized binding remained after 72 h at 4 degrees C. The affinities of solubilized (Kd = 11.4 +/- 0.5 nM) and membrane-bound receptors (Kd = 12.0 +/- 1.7 nM) were similar. The maximal number of binding sites/mg protein of solubilized receptors was 46 +/- 3% (n = 5) of membrane-bound receptors. Diuretics with a wide range of affinities had similar affinities for the solubilized and membrane-bound sites. Chloride inhibited and sodium stimulated the binding of [3H]metolazone to solubilized receptors, as they do with membrane-bound receptors. These studies demonstrate that, as judged by ligand binding, thiazide receptors can be solubilized in an active conformation and provide the basis for future purification and reconstitution.
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pubmed:grant |
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Apr
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pubmed:issn |
0006-3002
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
9
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pubmed:volume |
1052
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
119-22
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pubmed:dateRevised |
2007-11-14
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pubmed:meshHeading |
pubmed-meshheading:2322588-Animals,
pubmed-meshheading:2322588-Benzothiadiazines,
pubmed-meshheading:2322588-Binding, Competitive,
pubmed-meshheading:2322588-Binding Sites,
pubmed-meshheading:2322588-Cell Membrane,
pubmed-meshheading:2322588-Detergents,
pubmed-meshheading:2322588-Diuretics,
pubmed-meshheading:2322588-Kidney,
pubmed-meshheading:2322588-Kidney Tubules, Distal,
pubmed-meshheading:2322588-Kinetics,
pubmed-meshheading:2322588-Male,
pubmed-meshheading:2322588-Metolazone,
pubmed-meshheading:2322588-Phosphatidylcholines,
pubmed-meshheading:2322588-Protease Inhibitors,
pubmed-meshheading:2322588-Rats,
pubmed-meshheading:2322588-Rats, Inbred Strains,
pubmed-meshheading:2322588-Receptors, Drug,
pubmed-meshheading:2322588-Sodium Chloride Symporter Inhibitors,
pubmed-meshheading:2322588-Solubility
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pubmed:year |
1990
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pubmed:articleTitle |
Solubilization of thiazide diuretic receptors from rat kidney membranes.
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pubmed:affiliation |
Department of Medicine, University of California, San Diego, La Jolla 92093.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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