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Intercellular transfer of proteins as identified by stable isotope labeling of amino acids in cell culture.

Abstract
We tracked the extracellular fate of proteins of pulmonary origin using the technique of stable isotope labeling of amino acids in cell culture (SILAC) in cell-impermeable Transwell culture systems. We find that irradiation to murine lung and lung-derived cells induces their release of proteins that are capable of entering neighboring cells, including primary murine bone marrow cells as well as prostate cancer and hematopoietic cell lines. The functional classification of transferred proteins was broad and included transcription factors, mediators of basic cellular processes and components of the nucleosome remodeling and deacetylase complex, including metastasis associated protein 3 and retinoblastoma-binding protein 7. In further analysis we find that retinoblastoma-binding protein 7 is a transcriptional activator of E-cadherin and that its intercellular transfer leads to decreased gene expression of downstream targets such as N-cadherin and vimentin. SILAC-generated data sets offer a valuable tool to identify and validate potential paracrine networks that may impact relevant biologic processes associated with phenotypic and genotypic signatures of health and disease.
AuthorsMing Li, Jason M Aliotta, John M Asara, Qian Wu, Mark S Dooner, Lynne D Tucker, Alan Wells, Peter J Quesenberry, Bharat Ramratnam
JournalThe Journal of biological chemistry (J Biol Chem) Vol. 285 Issue 9 Pg. 6285-97 (Feb 26 2010) ISSN: 1083-351X [Electronic] United States
PMID20026604 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)
Chemical References
  • Amino Acids
  • Proteins
Topics
  • Amino Acids
  • Animals
  • Bone Marrow Cells (metabolism)
  • Cell Line
  • Cells, Cultured
  • Hematopoietic Stem Cells (metabolism)
  • Humans
  • Isotope Labeling (methods)
  • Lung (chemistry, cytology, radiation effects)
  • Male
  • Mice
  • Paracrine Communication (radiation effects)
  • Prostatic Neoplasms (metabolism)
  • Proteins (analysis, metabolism)
  • Proteomics (methods)

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