Expression of LY6D is induced at the surface of MCF10A cells by X‐ray irradiation

In order to identify membrane proteins whose expression is induced by X‐ray irradiation, we developed an antibody (Ab)‐directed strategy using a phage Ab library. X‐Ray‐irradiated cells were screened with a phage Ab library in the presence of a large excess of polyclonal Abs prepared against membrane proteins that are commonly present at the surface of both X‐ray‐irradiated and nonirradiated cells. After isolation of Ab that bound only to X‐ray‐irradiated cells, the antigen was identified using MS. Using this approach, we found that expression of LY6D is induced in MCF10A cells by X‐ray irradiation. The induction of LY6D expression is triggered through a pathway regulated by ATM, CHK2 and p53. This method is a new Ab‐directed proteomic strategy for analysis of membrane proteins, and is applicable to various biological phenomena in situations in which both target molecule‐expressing cells and nonexpressing cells are available. In order to identify membrane proteins whose expression is induced by X‐ray irradiation, we developed an antibody (Ab)‐directed strategy using a phage Ab library. Using this approach, we found that expression of LY6D is induced in MCF10A cells by X‐ray irradiation. This method is a new proteomic strategy for analysis of membrane proteins, and is applicable to various biological phenomena