Advertisement

Straightforward Protocol for Gel-Free Proteomic Analysis of Adipose Tissue

  • Yvonne Pasing
  • Armin Schniers
  • Terkel Hansen
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1788)

Abstract

After conducting systematic and quantitative comparisons of different sample preparation techniques regarding their capability to efficiently and reproducibly recover proteins from biopsies, we present here our superior protocol for extracting proteins from low amounts of adipose tissue. Adipose tissue as a matrix in bottom-up proteomics is challenging due to the extremely high lipid content.

The lysis buffer utilized contains the detergent sodium deoxycholate, which does not impair the activity of trypsin and therefore enables direct digestion without detergent removal steps. The resulting workflow is time saving, cost efficient, easy to perform, and it can also be applied to other hydrophobic samples.

Keywords

Adipose tissue proteome Detergents Hydrophobic samples Sample preparation Sodium deoxycholate Sodium laurate 

Notes

Acknowledgment

We would like to thank Prof. Rolf Jorde, Sayda Colnoe, Allan Didriksen, and Jack-Ansgar Bruun. Our work was supported by a grant from the North Norway Regional Health Authorities.

References

  1. 1.
    Galic S, Oakhill JS, Steinberg GR (2010) Adipose tissue as an endocrine organ. Mol Cell Endocrinol 316(2):129–139CrossRefGoogle Scholar
  2. 2.
    Matsuzawa Y (2006) Therapy insight: adipocytokines in metabolic syndrome and related cardiovascular disease. Nat Clin Pract Cardiovasc Med 3(1):35–42CrossRefGoogle Scholar
  3. 3.
    Peral B, Camafeita E, Fernandez-Real JM et al (2009) Tackling the human adipose tissue proteome to gain insight into obesity and related pathologies. Expert Rev Proteomics 6(4):353–361CrossRefGoogle Scholar
  4. 4.
    Pasing Y, Colnoe S, Hansen T (2017) Proteomics of hydrophobic samples: fast, robust and low-cost workflows for clinical approaches. Proteomics 17(6)Google Scholar
  5. 5.
    Schniers A, Anderssen E, Fenton CG et al. The proteome of ulcerative colitis in colon biopsies from adults—optimized sample preparation and comparison with healthy controls. Proteomics Clin Appl. Epub ahead of print. Doi:  https://doi.org/10.1002/prca.201700053
  6. 6.
    Lin Y, Huo L, Liu Z et al (2013) Sodium laurate, a novel protease- and mass spectrometry-compatible detergent for mass spectrometry-based membrane proteomics. PLoS One 8(3):e59779.  https://doi.org/10.1371/journal.pone.0059779 CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Zhou J, Zhou T, Cao R et al (2006) Evaluation of the application of sodium deoxycholate to proteomic analysis of rat hippocampal plasma membrane. J Proteome Res 5(10):2547–2553CrossRefGoogle Scholar
  8. 8.
    Leon IR, Schwammle V, Jensen ON et al (2013) Quantitative assessment of in-solution digestion efficiency identifies optimal protocols for unbiased protein analysis. Mol Cell Proteomics 12(10):2992–3005CrossRefGoogle Scholar
  9. 9.
    Lin Y, Wang K, Liu Z et al (2015) Enhanced SDC-assisted digestion coupled with lipid chromatography-tandem mass spectrometry for shotgun analysis of membrane proteome. J Chromatogr B 1002:144–151CrossRefGoogle Scholar
  10. 10.
    Fu SL, Li JL, Chen J et al (2015) Extraction and identification of membrane proteins from black widow spider eggs. Zool Res 36(4):248–254PubMedPubMedCentralGoogle Scholar
  11. 11.
    Masuda T, Tomita M, Ishihama Y (2008) Phase transfer surfactant-aided trypsin digestion for membrane proteome analysis. J Proteome Res 7(2):731–740CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York  2017

Authors and Affiliations

  1. 1.Tromsø Endocrine Research Group, Department of Clinical MedicineUiT – The Arctic University of NorwayTromsøNorway
  2. 2.Division of Internal MedicineUniversity Hospital of North NorwayTromsøNorway
  3. 3.Natural Products and Medicinal Chemistry Research Group, Department of PharmacyUiT – The Arctic University of NorwayTromsøNorway

Personalised recommendations