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Functionalized Peptidisc Scaffolds Enable Affinity Purification of the Membrane Proteome

ABSTRACT: We reconstituted the total E. coli membrane proteome into His-tagged peptidiscs. We then performed an affinity purification to enrich the bona fide membrane proteome away from soluble contaminants which co-sediment with cellular membranes after cell lysis. As a case study, we employ this method to survey changes to the membrane proteome caused by altered gene expression.

INSTRUMENT(S): impact II

ORGANISM(S): Escherichia Coli

SUBMITTER: David Rattray

LAB HEAD: Franck Van Hoa Duong

PROVIDER: PXD017242 | Pride | 2022-04-11

REPOSITORIES: Pride

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His-Tagged Peptidiscs Enable Affinity Purification of the Membrane Proteome for Downstream Mass Spectrometry Analysis.

Young John William JW Wason Irvinder Singh IS Zhao Zhiyu Z Rattray David G DG Foster Leonard J LJ Duong Van Hoa Franck F

Journal of proteome research 20200504 7

Characterization of the integral membrane proteome by mass spectrometry (MS) remains challenging due its high complexity and inherent insolubility. In a typical experiment, the cellular membranes are isolated, the proteins are solubilized and fractionated, and the detergent micelles are removed before MS analysis. Detergents are not compatible with mass spectrometry, however, and their removal from biological samples often results in reduced protein identification. As an alternative to detergent ...[more]

PMID: 32364744

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