ポスター発表
- 第3日 5月17日（木） ポスター会場
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3P-43 PDF
2D-DIGE法によるエクトドメインシェディングの解析
[Objective] Cell surface proteins digested at their extracellular domains and released from the surface (ectodomain shedding) are possibly associated with pathophysiology of various diseases. There has been no method to comprehensively detect these shedded proteins. Previously, we reported a method for detection of the proteins. In the current study, we tried to analyze the proteins by 2D-DIGE. [Methods] We used a human cervical carcinoma cell line of HeLa which were previously reported to express A Disintegrin And Metalloprotease 17 (ADAM17). Cell surface proteins of living Hela cells were labeled by Cy5 dye and then the cells were cultured in a medium containing TAPI-2 (ADAM17 inhibitor). As an internal control, cell surface proteins of which were labeled with Cy3 dye were cultured. Proteins in the cultured media were isolated and then were analyzed by 2D-DIGE. [Results] Many protein spots were visualized by 2D-DIGE. Multiple protein spots decreased their intensity by the treatment with TAPI-2. [Conclusion] We found that multiple protein spots were affected by TAPI-2. The affected protein would be released from the cell surface by ADAM17. This method is thought useful to comprehensively analyze cell surface proteins affected by the ectodomain shedding.