A method for the comprehensive proteomic analysis of membrane proteins - PubMed (original) (raw)

doi: 10.1038/nbt819. Epub 2003 Apr 14.

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• PMID: 12692561
• DOI: 10.1038/nbt819

A method for the comprehensive proteomic analysis of membrane proteins

Christine C Wu et al. Nat Biotechnol. 2003 May.

Abstract

We describe a method that allows for the concurrent proteomic analysis of both membrane and soluble proteins from complex membrane-containing samples. When coupled with multidimensional protein identification technology (MudPIT), this method results in (i) the identification of soluble and membrane proteins, (ii) the identification of post-translational modification sites on soluble and membrane proteins, and (iii) the characterization of membrane protein topology and relative localization of soluble proteins. Overlapping peptides produced from digestion with the robust nonspecific protease proteinase K facilitates the identification of covalent modifications (phosphorylation and methylation). High-pH treatment disrupts sealed membrane compartments without solubilizing or denaturing the lipid bilayer to allow mapping of the soluble domains of integral membrane proteins. Furthermore, coupling protease protection strategies to this method permits characterization of the relative sidedness of the hydrophilic domains of membrane proteins.

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Comment in

• Membrane proteins ride shotgun.
  Rabilloud T. Rabilloud T. Nat Biotechnol. 2003 May;21(5):508-10. doi: 10.1038/nbt0503-508. Nat Biotechnol. 2003. PMID: 12721575 No abstract available.

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