APols-Aided Protein Precipitation: A Rapid Method for Concentrating Proteins for Proteomic Analysis
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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.
KeywordsAmphipols Proteomics Protein precipitation Concentrating protein Mass spectrometry
D. F. acknowledges a Canada Research Chair in Proteomics and Systems Biology. Funding for this project was provided by NSERC-Canada. Z. N. acknowledges a postdoctoral scholarship from the CIHR Training Program in Neurodegenerative Lipidomics (TGF-96121). Z. N. would also like to acknowledge Dr. Jean-Luc POPOT for the valuable discussion and advices, and Alexandra Therese Star’s help for proofing.
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