Intercellular Transfer of Proteins as Identified by Stable Isotope Labeling of Amino Acids in Cell Culture

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...

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Bibliographic Details
Published in:The Journal of biological chemistry 2010-02, Vol.285 (9), p.6285-6297
Main Authors: Li, Ming, Aliotta, Jason M., Asara, John M., Wu, Qian, Dooner, Mark S., Tucker, Lynne D., Wells, Alan, Quesenberry, Peter J., Ramratnam, Bharat
Format: Article
Language:English
Subjects:
Amino Acids
Animals
Bone Marrow
Bone Marrow Cells - metabolism
Cancer
Cell Line
Cell/Cell-Cell Interaction
Cells, Cultured
Genomics and Proteomics
Hematopoietic Stem Cells - metabolism
Humans
Isotope Labeling - methods
Lung - chemistry
Lung - cytology
Lung - radiation effects
Male
Mice
Paracrine Communication - radiation effects
Prostatic Neoplasms - metabolism
Protein-DNA Interactions
Proteins - analysis
Proteins - metabolism
Proteomics
Proteomics - methods
Stem Cells
Transcription/Promoter
Transcription/Regulation
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Summary: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.
ISSN:0021-9258
1083-351X
1083-351X
DOI:10.1074/jbc.M109.057943