Comprehensive analysis of intracellular proteins on the cell surface for elucidation of their roles in tissue damage and repair.

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Although various intracellular proteins are released from damaged tissues, functional impact of these proteins in the extracellular space remains largely unknown. To characterize intracellular proteins potentially functioning in the extracellular space and affecting cellular responses, cell surface-associated proteins were selectively identified using a combination of cell surface labeling and mass spectrometry-based protein identification technology. We identified 454 proteins, including 405 putative intracellular proteins, on the surface of U937 cells. Besides a variety of high-abundance proteins, the protein subset included some of clinical markers, such as HMGB1. Out of the 405 proteins, 162 proteins were also found to be released from damaged HEK293 cells. Furthermore, a certain protein was confirmed to affect a signal transduction pathway in U937 cells.