The mammary epithelial cell secretome and its regulation by signal transduction pathways.
Extracellular proteins released by mammary epithelial cells are critical mediators of cell communication, proliferation, and organization, yet the actual spectrum of proteins released by any given cell (the secretome) is poorly characterized. To define the set of proteins secreted by human mammary epithelial cells (HMEC), we combined analytical and computational approaches to define a secretome protein set based upon probable biological significance. Analysis of HMEC-conditioned medium by liquid chromatography-mass spectrometry resulted in identification of 889 unique proteins, of which 151 were found to be specifically enriched in the extracellular compartment when compared with a database of proteins expressed in whole HMEC lysates. Additional high mass accuracy analysis revealed 36 proteins whose extracellular abundance increased after treatment with phorbol ester (PMA), a protein kinase C agonist and general secretagogue. Many of the PMA stimulated proteins have been reported to be aberrantly expressed in human cancers and appear to be coregulated as multigene clusters. By inhibiting PMA-mediated transactivation of the epidermal growth factor receptor (EGFR), a pathway critically required for normal HMEC function, we found that the secretion of specific matrix metalloproteases was also coordinately regulated through EGFR transactivation. This study demonstrates a tiered strategy by which extracellular proteins can be identified and progressively assigned to classes of increasing confidence and regulatory importance.
Jacobs
Jon M
JM
Systems Biology Program, Cell Biology and Biochemistry, Biological Separations and Mass Spectrometry, Computational Biology Groups, and Environmental Molecular Science Laboratory, Pacific Northwest National Laboratory, Richland, Washington 99352, USA.
Waters
Katrina M
KM
Kathmann
Loel E
LE
Camp
David G
DG
Wiley
H Steven
HS
Smith
Richard D
RD
Thrall
Brian D
BD
D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D004847:Epithelial Cells / Q000378:metabolism*; D066246:ErbB Receptors / Q000502:physiology; D005260:Female; D006801:Humans; D042361:Mammary Glands, Human / Q000166:cytology / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism*; D015203:Reproducibility of Results; D015398:Signal Transduction / Q000502:physiology*; D017550:Spectroscopy, Fourier Transform Infrared; D053719:Tandem Mass Spectrometry
D017077:Culture Media, Conditioned; D020543:Proteome; D066246:ErbB Receptors