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

Maiko Kurosawa, Anand D. Jeyasekharan, Eva Maria Surmann, Noboru Hashimoto, Vignesh Venkatraman, Gene Kurosawa, Koichi Furukawa, Ashok R. Venkitaraman, Yoshikazu Kurosawa

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Abstract

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.

Original languageEnglish
Pages (from-to)4479-4491
Number of pages13
JournalFEBS Journal
Volume279
Issue number24
DOIs
Publication statusPublished - 01-12-2012

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Kurosawa, M., Jeyasekharan, A. D., Surmann, E. M., Hashimoto, N., Venkatraman, V., Kurosawa, G., Furukawa, K., Venkitaraman, A. R., & Kurosawa, Y. (2012). Expression of LY6D is induced at the surface of MCF10A cells by X-ray irradiation. FEBS Journal, 279(24), 4479-4491. https://doi.org/10.1111/febs.12034