The influence of monovalent cations on trimeric G protein G(i)1α activity in HEK293 cells stably expressing DOR-G(i)1α (Cys(351)-Ile(351)) fusion protein
Language English Country Czech Republic Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
21401297
DOI
10.33549/physiolres.932096
PII: 932096
Knihovny.cz E-resources
- MeSH
- Metals, Alkali pharmacology MeSH
- Cysteine genetics MeSH
- HEK293 Cells MeSH
- Isoleucine genetics MeSH
- Cations, Monovalent MeSH
- Enkephalin, Leucine-2-Alanine pharmacology MeSH
- Humans MeSH
- Protein Multimerization MeSH
- GTP-Binding Protein alpha Subunits, Gi-Go genetics metabolism MeSH
- Receptors, Opioid, delta agonists genetics metabolism MeSH
- Recombinant Fusion Proteins genetics metabolism MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Metals, Alkali MeSH
- Cysteine MeSH
- Isoleucine MeSH
- Cations, Monovalent MeSH
- Enkephalin, Leucine-2-Alanine MeSH
- GTP-Binding Protein alpha Subunits, Gi-Go MeSH
- Receptors, Opioid, delta MeSH
- Recombinant Fusion Proteins MeSH
The effect of monovalent cations on trimeric G protein G(i)1α was measured at equimolar concentration of chloride anion in pertussis-toxin (PTX)-treated HEK293 cells stably expressing PTX-insensitive DOR- G(i)1α (Cys(351)-Ile(351)) fusion protein by high-affinity [(35)S]GTPgammaS binding assay. The high basal level of binding was detected in absence of DOR agonist and monovalent ions and this high level was inhibited with the order of: Na(+) > K(+) > Li(+). The first significant inhibition was detected at 1 mM NaCl. The inhibition by monovalent ions was reversed by increasing concentrations of DOR agonist DADLE. The maximum DADLE response was also highest for sodium and decreased in the order of: Na(+) > K(+) ~ Li(+). Our data indicate i) an inherently high activity of trimeric G protein G(i)1α when expressed within DOR- G(i)1α fusion protein and determined in the absence of monovalent cations, ii) preferential sensitivity of DOR- G(i)1alpha to sodium as far as maximum of agonist response is involved.
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