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

Fingerprint

Laminin Receptors
Laminin
Epitopes
Carrier Proteins
Monoclonal Antibodies
Embryonal Carcinoma Stem Cells
Cells
Cell Line
Clostridium
Clostridium perfringens
Collodion
Density Gradient Centrifugation
Centrifugation
Confocal microscopy
Kidney Neoplasms
Neuraminidase
Cell membranes
Galactose
Confocal Microscopy
Detergents

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Laminin binding protein, 34/67 laminin receptor, carries stage-specific embryonic antigen-4 epitope defined by monoclonal antibody Raft.2. / Katagiri, Yohko U.; Kiyokawa, Nobutaka; Nakamura, Kyoko; Takenouchi, Hisami; Taguchi, Tomoko; Okita, Hajime; Umezawa, Akihiro; Fujimoto, Junichiro.

In: Biochemical and Biophysical Research Communications, Vol. 332, No. 4, 15.07.2005, p. 1004-1011.

Research output: Contribution to journalArticle

Katagiri, Yohko U. ; Kiyokawa, Nobutaka ; Nakamura, Kyoko ; Takenouchi, Hisami ; Taguchi, Tomoko ; Okita, Hajime ; Umezawa, Akihiro ; Fujimoto, Junichiro. / Laminin binding protein, 34/67 laminin receptor, carries stage-specific embryonic antigen-4 epitope defined by monoclonal antibody Raft.2. In: Biochemical and Biophysical Research Communications. 2005 ; Vol. 332, No. 4. pp. 1004-1011.
@article{9c22ef1802d44e4da3a805d4b4ee4cec,
title = "Laminin binding protein, 34/67 laminin receptor, carries stage-specific embryonic antigen-4 epitope defined by monoclonal antibody Raft.2",
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.",
keywords = "Embryonal carcinoma, Laminin binding protein, Laminin receptor, MALDI-TOF MS, Raft.2, SialylGb5, SSEA-4",
author = "Katagiri, {Yohko U.} and Nobutaka Kiyokawa and Kyoko Nakamura and Hisami Takenouchi and Tomoko Taguchi and Hajime Okita and Akihiro Umezawa and Junichiro Fujimoto",
year = "2005",
month = "7",
day = "15",
doi = "10.1016/j.bbrc.2005.05.052",
language = "English",
volume = "332",
pages = "1004--1011",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "4",

}

TY - JOUR

T1 - Laminin binding protein, 34/67 laminin receptor, carries stage-specific embryonic antigen-4 epitope defined by monoclonal antibody Raft.2

AU - Katagiri, Yohko U.

AU - Kiyokawa, Nobutaka

AU - Nakamura, Kyoko

AU - Takenouchi, Hisami

AU - Taguchi, Tomoko

AU - Okita, Hajime

AU - Umezawa, Akihiro

AU - Fujimoto, Junichiro

PY - 2005/7/15

Y1 - 2005/7/15

N2 - 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.

AB - 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.

KW - Embryonal carcinoma

KW - Laminin binding protein

KW - Laminin receptor

KW - MALDI-TOF MS

KW - Raft.2

KW - SialylGb5

KW - SSEA-4

UR - http://www.scopus.com/inward/record.url?scp=20444403377&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=20444403377&partnerID=8YFLogxK

U2 - 10.1016/j.bbrc.2005.05.052

DO - 10.1016/j.bbrc.2005.05.052

M3 - Article

VL - 332

SP - 1004

EP - 1011

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 4

ER -