Journal
JOURNAL OF MEMBRANE BIOLOGY
Volume 247, Issue 9-10, Pages 941-947Publisher
SPRINGER
DOI: 10.1007/s00232-014-9668-6
Keywords
Amphipols; Proteomics; Protein precipitation; Concentrating protein; Mass spectrometry
Funding
- NSERC-Canada
- CIHR Training Program in Neurodegenerative Lipidomics [TGF-96121]
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Amphipols (APols) are a newly designed and milder class of detergent. They have been used primarily in protein structure analysis for membrane protein trapping and stabilization. We have recently demonstrated that APols can be used as an alternative detergent for proteome extraction and digestion, to achieve a One-stop single-tube workflow for proteomics. In this workflow, APols are removed by precipitation after protein digestion without depleting the digested peptides. Here, we took further advantage of this precipitation characteristic of APols to concentrate proteins from diluted samples. In contrast with tryptic peptides, a decrease in pH leads to the unbiased co-precipitation of APols with proteins, including globular hydrophilic proteins. We demonstrated that this precipitation is a combined effect of acid precipitation and the APols' protein interactions. Also, we have been able to demonstrate that APols-aided protein precipitation works well on diluted samples, such as secretome sample, and provides a rapid method for protein concentration.
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