Identification and classification of shuttle protein in and out of nuclei

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The nuclear protein prothymosin-α(ProTα), which possesses multi-function is an intrinsic inhibitor of necrosis and apoptosis. We have already revealed that ProTαis released from neurons and astrocytes on ischemic stress through non-vesicular manner and exerts a unique form of neuroprotection through an anti-necrotic mechanism. Previously, we found out that released ProTαonce again makes the transition to the nucleus in some cells. In this study, we comprehensively identified the ProTα-binding proteins in process of extracellular release and re-nuclear localization of ProTα, and performed protein-protein interaction analysis in in vitro and cell-based assay. Finally, we succeeded the identification of non-vesicular release involved proteins, ProTα-binding proteins in the nucleus and novel cell membrane receptor. In addition, we simulated variation of machineries involved in self-defensive role of ProTαby use of Cell Illustrator software, and obtained the similar result of experimental work.