APols-aided protein precipitation: a rapid method for concentrating proteins for proteomic analysis.
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.
Department of Biochemistry, Immunology and Microbiology, Ottawa Institute of Systems Biology, University of Ottawa, Ottawa, ON, Canada.
Zhibin
Z
Ning
Brett
B
Hawley
Deeptee
D
Seebun
Daniel
D
Figeys
D005590:Fractional Precipitation / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D011108:Polymers / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D020543:Proteome / Q000737:chemistry* / Q000302:isolation & purification*; D040901:Proteomics / Q000379:methods*; D012995:Solubility; D012996:Solutions; D013048:Specimen Handling; D013501:Surface-Active Agents / Q000737:chemistry*
D011108:Polymers; D011506:Proteins; D020543:Proteome; D012996:Solutions; D013501:Surface-Active Agents