Determination of membrane protein transporter oligomerization in native tissue using spatial fluorescence intensity fluctuation analysis.
Membrane transporter proteins exist in a complex dynamic equilibrium between various oligomeric states that include monomers, dimers, dimer of dimers and higher order oligomers. Given their sub-optical microscopic resolution size, the oligomerization state of membrane transporters is difficult to qu...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3338697?pdf=render |
| _version_ | 1818270883909730304 |
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| author | Mikhail Sergeev Antoine G Godin Liyo Kao Natalia Abuladze Paul W Wiseman Ira Kurtz |
| author_facet | Mikhail Sergeev Antoine G Godin Liyo Kao Natalia Abuladze Paul W Wiseman Ira Kurtz |
| author_sort | Mikhail Sergeev |
| collection | DOAJ |
| description | Membrane transporter proteins exist in a complex dynamic equilibrium between various oligomeric states that include monomers, dimers, dimer of dimers and higher order oligomers. Given their sub-optical microscopic resolution size, the oligomerization state of membrane transporters is difficult to quantify without requiring tissue disruption and indirect biochemical methods. Here we present the application of a fluorescence measurement technique which combines fluorescence image moment analysis and spatial intensity distribution analysis (SpIDA) to determine the oligomerization state of membrane proteins in situ. As a model system we analyzed the oligomeric state(s) of the electrogenic sodium bicarbonate cotransporter NBCe1-A in cultured cells and in rat kidney. The approaches that we describe offer for the first time the ability to investigate the oligomeric state of membrane transporter proteins in their native state. |
| first_indexed | 2024-12-12T21:17:22Z |
| format | Article |
| id | doaj.art-9b1161ce2b9c4ddabe2783eed8b5e5d8 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-12T21:17:22Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-9b1161ce2b9c4ddabe2783eed8b5e5d82022-12-22T00:11:42ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0174e3621510.1371/journal.pone.0036215Determination of membrane protein transporter oligomerization in native tissue using spatial fluorescence intensity fluctuation analysis.Mikhail SergeevAntoine G GodinLiyo KaoNatalia AbuladzePaul W WisemanIra KurtzMembrane transporter proteins exist in a complex dynamic equilibrium between various oligomeric states that include monomers, dimers, dimer of dimers and higher order oligomers. Given their sub-optical microscopic resolution size, the oligomerization state of membrane transporters is difficult to quantify without requiring tissue disruption and indirect biochemical methods. Here we present the application of a fluorescence measurement technique which combines fluorescence image moment analysis and spatial intensity distribution analysis (SpIDA) to determine the oligomerization state of membrane proteins in situ. As a model system we analyzed the oligomeric state(s) of the electrogenic sodium bicarbonate cotransporter NBCe1-A in cultured cells and in rat kidney. The approaches that we describe offer for the first time the ability to investigate the oligomeric state of membrane transporter proteins in their native state.http://europepmc.org/articles/PMC3338697?pdf=render |
| spellingShingle | Mikhail Sergeev Antoine G Godin Liyo Kao Natalia Abuladze Paul W Wiseman Ira Kurtz Determination of membrane protein transporter oligomerization in native tissue using spatial fluorescence intensity fluctuation analysis. PLoS ONE |
| title | Determination of membrane protein transporter oligomerization in native tissue using spatial fluorescence intensity fluctuation analysis. |
| title_full | Determination of membrane protein transporter oligomerization in native tissue using spatial fluorescence intensity fluctuation analysis. |
| title_fullStr | Determination of membrane protein transporter oligomerization in native tissue using spatial fluorescence intensity fluctuation analysis. |
| title_full_unstemmed | Determination of membrane protein transporter oligomerization in native tissue using spatial fluorescence intensity fluctuation analysis. |
| title_short | Determination of membrane protein transporter oligomerization in native tissue using spatial fluorescence intensity fluctuation analysis. |
| title_sort | determination of membrane protein transporter oligomerization in native tissue using spatial fluorescence intensity fluctuation analysis |
| url | http://europepmc.org/articles/PMC3338697?pdf=render |
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