Dominant portion of thyrotropin-releasing hormone receptor is excluded from lipid domains. Detergent-resistant and detergent-sensitive pools of TRH receptor and Gqalpha/G11alpha protein
Language English Country Great Britain, England Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
Grant support
Wellcome Trust - United Kingdom
PubMed
16091585
DOI
10.1093/jb/mvi114
PII: 138/2/111
Knihovny.cz E-resources
- MeSH
- Cell Membrane chemistry MeSH
- Cell Culture Techniques MeSH
- Centrifugation, Density Gradient MeSH
- Detergents MeSH
- Spectrometry, Fluorescence MeSH
- Immunoblotting MeSH
- Caveolin 1 MeSH
- Caveolins chemistry MeSH
- Humans MeSH
- Membrane Microdomains chemistry MeSH
- Octoxynol MeSH
- GTP-Binding Protein alpha Subunits, Gq-G11 analysis chemistry MeSH
- Radioligand Assay MeSH
- Receptors, Thyrotropin-Releasing Hormone analysis chemistry MeSH
- Solubility MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- CAV1 protein, human MeSH Browser
- Detergents MeSH
- Caveolin 1 MeSH
- Caveolins MeSH
- Octoxynol MeSH
- GTP-Binding Protein alpha Subunits, Gq-G11 MeSH
- Receptors, Thyrotropin-Releasing Hormone MeSH
Some G protein-coupled receptors might be spacially targetted to discrete domains within the plasma membrane. Here we assessed the localization in membrane domains of the epitope-tagged, fluorescent version of thyrotropin-releasing hormone receptor (VSV-TRH-R-GFP) expressed in HEK293 cells. Our comparison of three different methods of cell fractionation (detergent extraction, alkaline treatment/sonication and mechanical homogenization) indicated that the dominant portion of plasma membrane pool of the receptor was totally solubilized by Triton X-100 and its distribution was similar to that of transmembrane plasma membrane proteins (glycosylated and non-glycosylated forms of CD147, MHCI, CD29, CD44, transmembrane form of CD58, Tapa1 and Na,K-ATPase). As expected, caveolin and GPI-bound proteins CD55, CD59 and GPI-bound form of CD58 were preferentially localized in detergent-resistant membrane domains (DRMs). Trimeric G proteins G(q)alpha/G(11)alpha, G(i)alpha1/G(i)alpha2, G(s)alphaL/G(s)alphaS and Gbeta were distributed almost equally between detergent-resistant and detergent-solubilized pools. In contrast, VSV-TRH-R-GFP, Galpha, Gbeta and caveolin were localized massively only in low-density membrane fragments of plasma membranes, which were generated by alkaline treatment/sonication or by mechanical homogenization of cells. These data indicate that VSV-TRH-R-GFP as well as other transmembrane markers of plasma membranes are excluded from TX-100-resistant, caveolin-enriched membrane domains. Trimeric G protein G(q)alpha/G(11)alpha occurs in both DRMs and in the bulk of plasma membranes, which is totally solubilized by TX-100.
References provided by Crossref.org
Biochemical and physiological insights into TRH receptor-mediated signaling
The Role of Lipid Environment in Ganglioside GM1-Induced Amyloid β Aggregation
