Defining the membrane proteome of NK cells

Journal of Mass Spectrometry : JMS
Dhimankrishna GhoshJohn A Wilkins

Abstract

The present study was initiated to define the composition of the membrane proteome of the Natural Killer (NK) like cell line YTS. Isolated membranes were treated with reagents that have been reported to remove peripheral membrane proteins. Additional steps involving trifluoroethanol (TFE) were introduced in an effort to remove remaining nonintegral membrane proteins. This treatment resulted in the release of a subset of proteins without any apparent disruption of membrane integrity. The membranes were solubilized and digested with trypsin in 25% TFE. The resulting peptides were separated using an off-line two-dimensional reversed phase LC technique at alkaline and acidic pHs. Mass spectrometric analysis identified 1843 proteins with high confidence scores. On the basis of the presence of transmembrane regions or evidence of posttranslational modifications and prediction algorithms, approximately 40% of the identified proteins were predicted as plausible membrane proteins. The remaining species were largely involved in cellular processes and molecular functions that could be predicted to be transiently associated with membranes. The analytical approaches presented in this study offer robust generic methods for the identification...Continue Reading

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Citations

Jan 15, 2013·Molecular & Cellular Proteomics : MCP·Maxi ScheiterLothar Jänsch
Dec 24, 2020·International Journal of Molecular Sciences·Chang-Kyu HeoEun-Wie Cho
Nov 16, 2018·Journal of the American Chemical Society·Alma FujisawaItaru Hamachi

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