Method for Recovery and Immunoaffinity Enrichment of Membrane Proteins Illustrated with Metastatic Ovarian Cancer Tissues

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dc.contributor.authorSchneider L
dc.contributor.authorLikhte V
dc.contributor.authorWright W
dc.contributor.authorChu F
dc.contributor.authorCambron E
dc.contributor.authorBaldwin-Burnett A
dc.contributor.authorKrakow J
dc.contributor.authorSmejkal G
dc.date.accessioned2018-08-02T23:23:54Z
dc.date.available2018-08-02T23:23:54Z
dc.date.issued2012en
dc.date.updated2018-05-23T00:03:29Z
dc.description.abstractIntegral membrane proteins play key biological roles in cell signaling, transport, and pathogen invasion. However, quantitative clinical assays for this critical class of proteins remain elusive and are generally limited to serum-soluble extracellular fragments. Furthermore, classic proteomic approaches to membrane protein analysis typically involve proteolytic digestion of the soluble pieces, resulting in separation of intra- and extracellular segments and significant informational loss. In this paper, we describe the development of a new method for the quantitative extraction of intact integral membrane proteins (including GPCRs) from solid metastatic ovarian tumors using pressure cycling technology in combination with a new (ProteoSolve-TD) buffer system. This new extraction buffer is compatible with immunoaffinity methods (e.g., ELISA and immunoaffinity chromatography), as well as conventional proteomic techniques (e.g., 2D gels, western blots). We demonstrate near quantitative recovery of membrane proteins EDG2, EDG4, FASLG, KDR, and LAMP-3 by western blots. We have also adapted commercial ELISAs for serum-soluble membrane protein fragments (e.g., sVEGFR2) to measure the tissue titers of their transmembrane progenitors. Finally, we demonstrate the compatibility of the new buffers with immunoaffinity enrichment/mass spectrometric characterization of tissue proteins.en
dc.identifier.doihttps://doi.org/10.1155/2012/838630
dc.identifier.issn2090-2166
dc.identifier.issn2090-2174
dc.identifier.urihttp://hdl.handle.net/10092/15718
dc.language.isoen
dc.rightsCopyright © 2012 Luke V. Schneider et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.en
dc.subject.anzsrcField of Research::11 - Medical and Health Sciences::1112 - Oncology and Carcinogenesis::111201 - Cancer Cell Biologyen
dc.subject.anzsrcFields of Research::32 - Biomedical and clinical sciences::3208 - Medical physiology::320801 - Cell physiologyen
dc.subject.anzsrcFields of Research::32 - Biomedical and clinical sciences::3205 - Medical biochemistry and metabolomics::320506 - Medical biochemistry - proteins and peptides (incl. medical proteomics)en
dc.titleMethod for Recovery and Immunoaffinity Enrichment of Membrane Proteins Illustrated with Metastatic Ovarian Cancer Tissuesen
dc.typeJournal Articleen
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