Linked Life Data

8813603

Source:http://linkedlifedata.com/resource/pubmed/id/8813603

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1-3
pubmed:dateCreated
1997-2-12
pubmed:abstractText
Rat liver membranes (crude P2 membranes) were solubilized in 10 mM Tris-HCl, pH 7.4 containing 7 mM 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS). The soluble fraction was designated the Extract 1. The 105,000 x g pellet was washed once, and then extracted a second time (Extract 2). The various resulting fractions were assayed for sigma (sigma) binding characteristics, using [3H](+)-pentazocine to label sigma 1 sites and [3H]1,3-di-o-tolylguanidine (DTG) in the presence of 1 microM dextrallorphan to label sigma 2 sites. Both of the extracts and resultant pellets (Pellet 1 and Pellet 2) contained sigma 1 and sigma 2 receptors, as indicated by the pharmacological profiles upon competition studies. The Kd and Bmax values for sigma 1 activity in the original P2 membranes were 8.3 +/- 0.73 nM and 5333 +/- 572 fmol/mg protein; Kd and Bmax for sigma 2 activity was 19 +/- 0.17 nM and 9190 +/- 800 fmol/mg protein. There were no changes in the radioligand Kd values of the two sites in the subsequent soluble and particulate fractions. However, while the sigma 1 and sigma 2 Bmax values in extracts and pellets were generally on the same order as those of P2 membranes, the actual sigma 2 to sigma 1 Bmax ratio varied markedly across the fractions. The ratio of sigma 2/ sigma 1 binding in Extract 1 and Extract 2 was 0.86 and 0.68, respectively, compared to a ratio of 1.7 in the original P2. However, the ratio in Pellet 2 was 3.8, twice that of the original P2 membranes. Furthermore, the Bmax value for sigma 1 sites in Pellet 2 did not change, whereas the sigma 2 Bmax increased 1.8 fold relative to the original P2 membranes. The changes in sigma 2/ sigma 1 binding ratio in extracts were observed using two different assay methods for soluble receptors (retention on polyethyleneimine-coated filters and polyethylene glycol precipitation) and is therefore not an artifact of assay procedure. These data suggest that, relative to sigma 1 receptors, sigma 2 receptors are more resistant to solubilization and become somewhat enriched in the particulate fractions. This supports the notion that sigma 1 and sigma 2 receptors are distinct macromolecules and may indicate different modes of association with the cell membrane.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
May
pubmed:issn
0014-2999
pubmed:author
pubmed:issnType
Print
pubmed:day
23
pubmed:volume
304
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
201-10
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed:year
1996
pubmed:articleTitle
Differential solubilization of rat liver sigma 1 and sigma 2 receptors: retention of sigma 2 sites in particulate fractions.
pubmed:affiliation
Unit on Receptor Biochemistry and Pharmacology, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD 20892, USA.
pubmed:publicationType
Journal Article, In Vitro