Laminin binding protein, 34/67 laminin receptor, carries stage-specific embryonic antigen-4 epitope defined by monoclonal antibody Raft.2

Yohko U. Katagiri, Nobutaka Kiyokawa, Kyoko Nakamura, Hisami Takenouchi, Tomoko Taguchi, Hajime Okita, Akihiro Umezawa, Junichiro Fujimoto

Research output: Contribution to journalArticle

12 Citations (Scopus)

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 [14C]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.

Original languageEnglish
Pages (from-to)1004-1011
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume332
Issue number4
DOIs
Publication statusPublished - 2005 Jul 15
Externally publishedYes

Keywords

  • Embryonal carcinoma
  • Laminin binding protein
  • Laminin receptor
  • MALDI-TOF MS
  • Raft.2
  • SSEA-4
  • SialylGb5

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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