Plasma membrane (PM) proteins contribute to the identity of a cell, mediate contact and communication and account for more than two thirds of known drug targets1-8. In the past years several protocols for the proteomic profiling of PM proteins have been described. Nevertheless, comparative analyses have mainly focused on different variations of one approach9-11. We compared sulfo-NHS-SS-biotinylation, aminooxy-biotinylation and surface coating with silica beads to isolate PM proteins for subsequent analysis by one-dimensional gel-free liquid chromatography mass spectrometry. Absolute and relative numbers of PM proteins and reproducibility parameters on a qualitative and quantitative level were assessed. Sulfo-NHS-SS-biotinylation outperformed aminooxy-biotinylation and surface coating using silica beads for most of the monitored criteria. We further simplified this procedure by a competitive biotin elution strategy achieving an average PM annotated protein fraction of 54 % (347 proteins). Computational analysis using additional databases and prediction tools revealed that in total over 90 % of the purified proteins were associated with the PM, mostly as interactors. The modified sulfo-NHS-SS-biotinylation protocol was validated by tracking changes in the plasma membrane proteome composition induced by genetic alteration and drug treatment. GPI-anchored proteins were depleted in PM purifications from cells deficient in the GPI transamidase component PIGS; and treatment of cells with tunicamycin significantly reduced the abundance of N-glycoproteins in surface purifications.
Pharmacology, Toxicology and Pharmaceutical Science
6
5%
Medicine and Dentistry
5
4%
Other
8
7%
Unknown
35
30%
Attention Score in Context
Attention Score in Context
This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 13 January 2016.
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#20,300,248
of 22,837,982 outputs
Outputs from Journal of Proteome Research
#5,702
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Outputs of similar age
#331,825
of 395,128 outputs
Outputs of similar age from Journal of Proteome Research
#86
of 97 outputs
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