Display options
Cite
Schneider LV, Likhte V, Wright WH, et al. Method for recovery and immunoaffinity enrichment of membrane proteins illustrated with metastatic ovarian cancer tissues. Int J Proteomics. 2012;2012:838630doi: 10.1155/2012/838630.
Schneider, L. V., Likhte, V., Wright, W. H., Chu, F., Cambron, E., Baldwin-Burnett, A., Krakow, J., & Smejkal, G. B. (2012). Method for recovery and immunoaffinity enrichment of membrane proteins illustrated with metastatic ovarian cancer tissues. International journal of proteomics, 2012838630. https://doi.org/10.1155/2012/838630
Schneider, Luke V, et al. "Method for recovery and immunoaffinity enrichment of membrane proteins illustrated with metastatic ovarian cancer tissues." International journal of proteomics vol. 2012 (2012): 838630. doi: https://doi.org/10.1155/2012/838630
Schneider LV, Likhte V, Wright WH, Chu F, Cambron E, Baldwin-Burnett A, Krakow J, Smejkal GB. Method for recovery and immunoaffinity enrichment of membrane proteins illustrated with metastatic ovarian cancer tissues. Int J Proteomics. 2012;2012:838630. doi: 10.1155/2012/838630. Epub 2012 Jul 12. PMID: 22919487; PMCID: PMC3418661.
Copy Download .nbib Format: NLM
Share it on

Int J Proteomics. 2012;2012:838630. doi: 10.1155/2012/838630. Epub 2012 Jul 12.

Method for recovery and immunoaffinity enrichment of membrane proteins illustrated with metastatic ovarian cancer tissues.

International journal of proteomics

Luke V Schneider, Varsha Likhte, William H Wright, Frances Chu, Emma Cambron, Anne Baldwin-Burnett, Jessica Krakow, Gary B Smejkal

Affiliations

  1. Target Discovery, Inc., 4030 Fabian Way, Palo Alto, CA 94030, USA.

PMID: 22919487 PMCID: PMC3418661 DOI: 10.1155/2012/838630

Abstract

Integral 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.

References

  1. Protein Sci. 2003 Feb;12(2):218-27 - PubMed
  2. Contraception. 2004 Aug;70(2):141-7 - PubMed
  3. Mol Cell Proteomics. 2003 Nov;2(11):1234-43 - PubMed
  4. Electrophoresis. 1999 Dec;20(18):3551-67 - PubMed
  5. Breast J. 2008 Jan-Feb;14(1):33-8 - PubMed
  6. Rapid Commun Mass Spectrom. 2007;21(6):837-45 - PubMed
  7. Oncol Res. 2007;16(9):445-51 - PubMed
  8. Drug Discov Today. 2005 Mar 1;10(5):353-63 - PubMed
  9. Proteomics. 2007 May;7(10):1654-63 - PubMed
  10. Anal Biochem. 2004 Oct 1;333(1):174-81 - PubMed
  11. J Chromatogr B Analyt Technol Biomed Life Sci. 2009 May 1;877(13):1229-39 - PubMed
  12. Methods Mol Biol. 2008;424:139-46 - PubMed
  13. Electrophoresis. 1997 Mar-Apr;18(3-4):533-7 - PubMed
  14. J Mass Spectrom. 2003 Aug;38(8):809-16 - PubMed
  15. Biochem J. 1999 Aug 15;342 ( Pt 1):119-23 - PubMed
  16. Biotechniques. 2007 Oct;43(4):427-30, 432, 434 passim - PubMed
  17. Anal Chem. 1995 Apr 1;67(7):1153-8 - PubMed
  18. J Clin Microbiol. 1995 Feb;33(2):419-27 - PubMed
  19. J Mol Biol. 1999 Oct 22;293(2):367-79 - PubMed
  20. Mol Cell Biol. 1999 Mar;19(3):1720-30 - PubMed
  21. Clin Chem. 2003 Jul;49(7):1041-4 - PubMed
  22. Am J Clin Pathol. 2004 Jun;121 Suppl:S33-49 - PubMed
  23. Clin Chem. 2002 Dec;48(12):2105-14 - PubMed
  24. J Proteome Res. 2006 Mar;5(3):695-700 - PubMed
  25. Expert Rev Proteomics. 2004 Dec;1(4):381-7 - PubMed
  26. Biomark Insights. 2007 Feb 07;1:1-48 - PubMed
  27. Mol Cell Proteomics. 2004 Apr;3(4):311-26 - PubMed
  28. Rapid Commun Mass Spectrom. 2008;22(6):905-8 - PubMed
  29. J Biol Chem. 1984 Nov 10;259(21):13241-5 - PubMed
  30. Protein Sci. 1994 Oct;3(10):1712-25 - PubMed
  31. IDrugs. 2009 May;12(5):302-8 - PubMed
  32. J Virol. 1984 Aug;51(2):340-5 - PubMed
  33. J Proteome Res. 2004 Mar-Apr;3(2):235-44 - PubMed
  34. Mol Cell Proteomics. 2004 Dec;3(12):1154-69 - PubMed
  35. Nat Rev Drug Discov. 2006 Dec;5(12):993-6 - PubMed
  36. Nat Biotechnol. 1999 Oct;17(10):994-9 - PubMed
  37. Expert Rev Proteomics. 2004 Dec;1(4):421-31 - PubMed
  38. Nat Rev Drug Discov. 2002 Sep;1(9):727-30 - PubMed
  39. Electrophoresis. 2004 Aug;25(15):2511-9 - PubMed
  40. J Proteome Res. 2008 Feb;7(2):659-65 - PubMed
  41. Cancer Res. 1993 Feb 15;53(4):747-54 - PubMed
  42. Biochem Int. 1986 Dec;13(6):927-32 - PubMed
  43. Anal Biochem. 1981 Apr;112(2):195-203 - PubMed
  44. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3466-70 - PubMed
  45. Science. 2000 Mar 17;287(5460):1960-4 - PubMed
  46. Proteomics. 2006 Oct;6(19):5269-77 - PubMed

Publication Types