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The Journal of Membrane Biology

, Volume 247, Issue 9–10, pp 941–947 | Cite as

APols-Aided Protein Precipitation: A Rapid Method for Concentrating Proteins for Proteomic Analysis

  • Zhibin Ning
  • Brett Hawley
  • Deeptee Seebun
  • Daniel FigeysEmail author
Article

Abstract

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.

Keywords

Amphipols Proteomics Protein precipitation Concentrating protein Mass spectrometry 

Notes

Acknowledgments

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.

Supplementary material

232_2014_9668_MOESM1_ESM.docx (957 kb)
Supplementary material 1 (DOCX 956 kb)
232_2014_9668_MOESM2_ESM.xlsx (327 kb)
Supplementary material 2 (XLSX 327 kb)
232_2014_9668_MOESM3_ESM.xlsx (693 kb)
Supplementary material 3 (XLSX 692 kb)

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Authors and Affiliations

  • Zhibin Ning
    • 1
  • Brett Hawley
    • 1
  • Deeptee Seebun
    • 1
  • Daniel Figeys
    • 1
    Email author
  1. 1.Department of Biochemistry, Immunology and Microbiology, Ottawa Institute of Systems BiologyUniversity of OttawaOttawaCanada

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