Laminin binding protein, 34/67 laminin receptor, carries stage-specific embryonic antigen-4 epitope defined by monoclonal antibody Raft.2 |
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Authors: | Katagiri Yohko U Kiyokawa Nobutaka Nakamura Kyoko Takenouchi Hisami Taguchi Tomoko Okita Hajime Umezawa Akihiro Fujimoto Junichiro |
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Institution: | Department of Developmental Biology, National Research Institute for Child Health and Development, Setagaya-ku, Tokyo 157-8535, Japan. kata@nch.go.jp |
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Abstract: | We previously produced monoclonal antibodies against the detergent-insoluble microdomain, i.e., the raft microdomain, of the human renal cancer cell line ACHN. Raft.2, one of these monoclonal antibodies, recognizes sialosyl globopentaosylceramide, which has the stage-specific embryonic antigen (SSEA)-4 epitope. Although the mouse embryonal carcinoma (EC) cell line F9 does not express SSEA-4, some F9 cells stained with Raft.2. Western analysis and matrix-assisted laser desorption ionization-time of flight mass spectrometry identified the Raft.2 binding molecule as laminin binding protein (LBP), i.e., 34/67 laminin receptor. Weak acid treatment or digestion with Clostridium perfringens sialidase reduced Raft.2 binding to LBP on nitrocellulose sheets and (14)C]galactose was incorporated into LBP, indicating LBP to have a sialylated carbohydrate moiety. Subcellular localization analysis by sucrose density-gradient centrifugation and examination by confocal microscopy revealed LBP to be localized on the outer surface of the plasma membrane. An SSEA-4-positive human EC cell line, NCR-G3 cells, also expressed Raft.2-binding LBP. |
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Keywords: | SSEA-4 SialylGb5 Embryonal carcinoma Laminin binding protein Laminin receptor Raft 2 MALDI-TOF MS |
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