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Large-Scale Analysis of Breast Cancer-Related Conformational Changes in Proteins Using Limited Proteolysis.

Publication ,  Journal Article
Liu, F; Fitzgerald, MC
Published in: Journal of proteome research
December 2016

Conformational changes in proteins can lead to disease. Thus, methods for identifying conformational changes in proteins can further improve our understanding and facilitate detection of disease states. Here we combine limited proteolysis (LiP) with Stable Isotope Labeling with Amino Acids in Cell Culture (SILAC) to characterize breast cancer-related conformational changes in proteins on the proteomic scale. Studied here are the conformational properties of proteins in two cell culture models of breast cancer, including the MCF-10A and MCF-7 cell lines. The SILAC-LiP approach described here identified ∼200 proteins with cell-line-dependent conformational changes, as determined by their differential susceptibility to proteolytic digestion using the nonspecific protease, proteinase K. The protease susceptibility profiles of the proteins in these cell lines were compared to thermodynamic stability and expression level profiles previously generated for proteins in these same breast cancer cell lines. The comparisons revealed that there was little overlap between the proteins with protease susceptibility changes and the proteins with thermodynamic stability and/or expression level changes. Thus, the large-scale conformational analysis described here provides unique insight into the molecular basis of the breast cancer phenotypes in this study.

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Published In

Journal of proteome research

DOI

EISSN

1535-3907

ISSN

1535-3893

Publication Date

December 2016

Volume

15

Issue

12

Start / End Page

4666 / 4674

Related Subject Headings

  • Proteolysis
  • Protein Stability
  • Protein Conformation
  • Peptide Hydrolases
  • Neoplasm Proteins
  • MCF-7 Cells
  • Humans
  • Female
  • Cell Line, Tumor
  • Breast Neoplasms
 

Citation

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Chicago
ICMJE
MLA
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Liu, F., & Fitzgerald, M. C. (2016). Large-Scale Analysis of Breast Cancer-Related Conformational Changes in Proteins Using Limited Proteolysis. Journal of Proteome Research, 15(12), 4666–4674. https://doi.org/10.1021/acs.jproteome.6b00755
Liu, Fang, and Michael C. Fitzgerald. “Large-Scale Analysis of Breast Cancer-Related Conformational Changes in Proteins Using Limited Proteolysis.Journal of Proteome Research 15, no. 12 (December 2016): 4666–74. https://doi.org/10.1021/acs.jproteome.6b00755.
Liu F, Fitzgerald MC. Large-Scale Analysis of Breast Cancer-Related Conformational Changes in Proteins Using Limited Proteolysis. Journal of proteome research. 2016 Dec;15(12):4666–74.
Liu, Fang, and Michael C. Fitzgerald. “Large-Scale Analysis of Breast Cancer-Related Conformational Changes in Proteins Using Limited Proteolysis.Journal of Proteome Research, vol. 15, no. 12, Dec. 2016, pp. 4666–74. Epmc, doi:10.1021/acs.jproteome.6b00755.
Liu F, Fitzgerald MC. Large-Scale Analysis of Breast Cancer-Related Conformational Changes in Proteins Using Limited Proteolysis. Journal of proteome research. 2016 Dec;15(12):4666–4674.
Journal cover image

Published In

Journal of proteome research

DOI

EISSN

1535-3907

ISSN

1535-3893

Publication Date

December 2016

Volume

15

Issue

12

Start / End Page

4666 / 4674

Related Subject Headings

  • Proteolysis
  • Protein Stability
  • Protein Conformation
  • Peptide Hydrolases
  • Neoplasm Proteins
  • MCF-7 Cells
  • Humans
  • Female
  • Cell Line, Tumor
  • Breast Neoplasms