Skip to main content
Journal cover image

Comparative Analysis of Mass-Spectrometry-Based Proteomic Methods for Protein Target Discovery Using a One-Pot Approach.

Publication ,  Journal Article
Cabrera, A; Wiebelhaus, N; Quan, B; Ma, R; Meng, H; Fitzgerald, MC
Published in: Journal of the American Society for Mass Spectrometry
February 2020

Recently, several mass-spectrometry- and protein-denaturation-based proteomic methods have been developed to facilitate protein target discovery efforts in drug mode-of-action studies. These methods, which include the stability of proteins from rates of oxidation (SPROX), pulse proteolysis (PP), chemical denaturation and protein precipitation (CPP), and thermal proteome profiling (TPP) techniques, have been used in an increasing number of applications in recent years. However, while the advantages and disadvantages to using these different techniques have been reviewed, the analytical characteristics of these methods have not been directly compared. Reported here is such a direct comparison using the well-studied immunosuppressive drug, cyclosporine A (CsA), and the proteins in a yeast cell lysate. Also described is a one-pot strategy that can be utilized with each technique to streamline data acquisition and analysis. We find that there are benefits to utilizing all four strategies for protein target discovery including increased proteomic coverage and reduced false positive rates that approach 0%. Moreover, the one-pot strategy described here makes such an experiment feasible, because of the 10-fold reduction in reagent costs and instrument time it affords.

Duke Scholars

Altmetric Attention Stats
Dimensions Citation Stats

Published In

Journal of the American Society for Mass Spectrometry

DOI

EISSN

1879-1123

ISSN

1044-0305

Publication Date

February 2020

Volume

31

Issue

2

Start / End Page

217 / 226

Related Subject Headings

  • Proteomics
  • Proteome
  • Proteolysis
  • Protein Stability
  • Protein Denaturation
  • Oxidation-Reduction
  • Mass Spectrometry
  • Fungal Proteins
  • Drug Development
  • Cyclosporine
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Cabrera, A., Wiebelhaus, N., Quan, B., Ma, R., Meng, H., & Fitzgerald, M. C. (2020). Comparative Analysis of Mass-Spectrometry-Based Proteomic Methods for Protein Target Discovery Using a One-Pot Approach. Journal of the American Society for Mass Spectrometry, 31(2), 217–226. https://doi.org/10.1021/jasms.9b00041
Cabrera, Aurora, Nancy Wiebelhaus, Baiyi Quan, Renze Ma, He Meng, and Michael C. Fitzgerald. “Comparative Analysis of Mass-Spectrometry-Based Proteomic Methods for Protein Target Discovery Using a One-Pot Approach.Journal of the American Society for Mass Spectrometry 31, no. 2 (February 2020): 217–26. https://doi.org/10.1021/jasms.9b00041.
Cabrera A, Wiebelhaus N, Quan B, Ma R, Meng H, Fitzgerald MC. Comparative Analysis of Mass-Spectrometry-Based Proteomic Methods for Protein Target Discovery Using a One-Pot Approach. Journal of the American Society for Mass Spectrometry. 2020 Feb;31(2):217–26.
Cabrera, Aurora, et al. “Comparative Analysis of Mass-Spectrometry-Based Proteomic Methods for Protein Target Discovery Using a One-Pot Approach.Journal of the American Society for Mass Spectrometry, vol. 31, no. 2, Feb. 2020, pp. 217–26. Epmc, doi:10.1021/jasms.9b00041.
Cabrera A, Wiebelhaus N, Quan B, Ma R, Meng H, Fitzgerald MC. Comparative Analysis of Mass-Spectrometry-Based Proteomic Methods for Protein Target Discovery Using a One-Pot Approach. Journal of the American Society for Mass Spectrometry. 2020 Feb;31(2):217–226.
Journal cover image

Published In

Journal of the American Society for Mass Spectrometry

DOI

EISSN

1879-1123

ISSN

1044-0305

Publication Date

February 2020

Volume

31

Issue

2

Start / End Page

217 / 226

Related Subject Headings

  • Proteomics
  • Proteome
  • Proteolysis
  • Protein Stability
  • Protein Denaturation
  • Oxidation-Reduction
  • Mass Spectrometry
  • Fungal Proteins
  • Drug Development
  • Cyclosporine