Propolypeptide of von Willebrand factor is a novel ligand for very late antigen-4 integrin

Takashi Isobe, Tetsuya Hisaoka, Akira Shimizu, Mitsuhiro Okuno, Saburo Aimoto, Yoshikazu Takada, Yuji Saito, Junichi Takagi

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

We have previously reported that propolypeptide of von Willebrand factor (pp-vWF) promotes melanoma cell adhesion in a β1 integrin-dependent manner. In this report, we identified the α subunit of the cell adhesion receptor for pp-vWF as α4. Human leukemia cell lines that express α4β1 integrin (very late antigen-4, VLA-4), but not cell lines which lack VLA-4, attached well to pp-vWF substrate and these adhesions were completely inhibited by anti-α4 integrin monoclonal antibody HP2/1. Adhesion of mouse melanoma expressing α4 integrin was also inhibited by anti-mouse α4 mAb PS/2. Furthermore, transfection of human α4 cDNA into α4-Chinese hamster ovary cells resulted in an acquisition of adhesive activity to pp-vWF, indicating that pp-vWF is a ligand for VLA-4 integrin. Using a recombinant fragment of pp-vWF, the cell attachment site was shown to be located within amino acid residues 376-455 of pp-vWF. A series of synthetic peptides covering this region were tested for the ability to promote cell attachment and a 15- residue peptide designated T2-15 (DCQDHSF-SIVIETVQ, residues numbered 395- 409) promoted VLA-4 dependent cell adhesion. The peptide was also capable of inhibiting cell adhesion to pp-vWF, suggesting that this sequence represents the cell attachment site. By affinity chromatography using peptide T2-15- Sepharose, it was found that α4β1 integrin complex from extracts of surface iodinated B16 cells specifically bound to the peptide. These results strongly suggest that pp-vWF is a novel physiological ligand for VLA-4.

Original languageEnglish (US)
Pages (from-to)8447-8453
Number of pages7
JournalJournal of Biological Chemistry
Volume272
Issue number13
DOIs
StatePublished - Mar 28 1997
Externally publishedYes

Fingerprint

Integrin alpha4beta1
Integrins
Ligands
Cell adhesion
Cell Adhesion
Peptides
Cells
Melanoma
Adhesion
Affinity chromatography
von Willebrand factor propolypeptide
Cell Line
Cricetulus
Complex Mixtures
Affinity Chromatography
Adhesives
Sepharose
Transfection
Ovary
Leukemia

ASJC Scopus subject areas

  • Biochemistry

Cite this

Propolypeptide of von Willebrand factor is a novel ligand for very late antigen-4 integrin. / Isobe, Takashi; Hisaoka, Tetsuya; Shimizu, Akira; Okuno, Mitsuhiro; Aimoto, Saburo; Takada, Yoshikazu; Saito, Yuji; Takagi, Junichi.

In: Journal of Biological Chemistry, Vol. 272, No. 13, 28.03.1997, p. 8447-8453.

Research output: Contribution to journalArticle

Isobe, T, Hisaoka, T, Shimizu, A, Okuno, M, Aimoto, S, Takada, Y, Saito, Y & Takagi, J 1997, 'Propolypeptide of von Willebrand factor is a novel ligand for very late antigen-4 integrin', Journal of Biological Chemistry, vol. 272, no. 13, pp. 8447-8453. https://doi.org/10.1074/jbc.272.13.8447
Isobe, Takashi ; Hisaoka, Tetsuya ; Shimizu, Akira ; Okuno, Mitsuhiro ; Aimoto, Saburo ; Takada, Yoshikazu ; Saito, Yuji ; Takagi, Junichi. / Propolypeptide of von Willebrand factor is a novel ligand for very late antigen-4 integrin. In: Journal of Biological Chemistry. 1997 ; Vol. 272, No. 13. pp. 8447-8453.
@article{a18c5245b2e3471c9653a99bec9224ed,
title = "Propolypeptide of von Willebrand factor is a novel ligand for very late antigen-4 integrin",
abstract = "We have previously reported that propolypeptide of von Willebrand factor (pp-vWF) promotes melanoma cell adhesion in a β1 integrin-dependent manner. In this report, we identified the α subunit of the cell adhesion receptor for pp-vWF as α4. Human leukemia cell lines that express α4β1 integrin (very late antigen-4, VLA-4), but not cell lines which lack VLA-4, attached well to pp-vWF substrate and these adhesions were completely inhibited by anti-α4 integrin monoclonal antibody HP2/1. Adhesion of mouse melanoma expressing α4 integrin was also inhibited by anti-mouse α4 mAb PS/2. Furthermore, transfection of human α4 cDNA into α4-Chinese hamster ovary cells resulted in an acquisition of adhesive activity to pp-vWF, indicating that pp-vWF is a ligand for VLA-4 integrin. Using a recombinant fragment of pp-vWF, the cell attachment site was shown to be located within amino acid residues 376-455 of pp-vWF. A series of synthetic peptides covering this region were tested for the ability to promote cell attachment and a 15- residue peptide designated T2-15 (DCQDHSF-SIVIETVQ, residues numbered 395- 409) promoted VLA-4 dependent cell adhesion. The peptide was also capable of inhibiting cell adhesion to pp-vWF, suggesting that this sequence represents the cell attachment site. By affinity chromatography using peptide T2-15- Sepharose, it was found that α4β1 integrin complex from extracts of surface iodinated B16 cells specifically bound to the peptide. These results strongly suggest that pp-vWF is a novel physiological ligand for VLA-4.",
author = "Takashi Isobe and Tetsuya Hisaoka and Akira Shimizu and Mitsuhiro Okuno and Saburo Aimoto and Yoshikazu Takada and Yuji Saito and Junichi Takagi",
year = "1997",
month = "3",
day = "28",
doi = "10.1074/jbc.272.13.8447",
language = "English (US)",
volume = "272",
pages = "8447--8453",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "13",

}

TY - JOUR

T1 - Propolypeptide of von Willebrand factor is a novel ligand for very late antigen-4 integrin

AU - Isobe, Takashi

AU - Hisaoka, Tetsuya

AU - Shimizu, Akira

AU - Okuno, Mitsuhiro

AU - Aimoto, Saburo

AU - Takada, Yoshikazu

AU - Saito, Yuji

AU - Takagi, Junichi

PY - 1997/3/28

Y1 - 1997/3/28

N2 - We have previously reported that propolypeptide of von Willebrand factor (pp-vWF) promotes melanoma cell adhesion in a β1 integrin-dependent manner. In this report, we identified the α subunit of the cell adhesion receptor for pp-vWF as α4. Human leukemia cell lines that express α4β1 integrin (very late antigen-4, VLA-4), but not cell lines which lack VLA-4, attached well to pp-vWF substrate and these adhesions were completely inhibited by anti-α4 integrin monoclonal antibody HP2/1. Adhesion of mouse melanoma expressing α4 integrin was also inhibited by anti-mouse α4 mAb PS/2. Furthermore, transfection of human α4 cDNA into α4-Chinese hamster ovary cells resulted in an acquisition of adhesive activity to pp-vWF, indicating that pp-vWF is a ligand for VLA-4 integrin. Using a recombinant fragment of pp-vWF, the cell attachment site was shown to be located within amino acid residues 376-455 of pp-vWF. A series of synthetic peptides covering this region were tested for the ability to promote cell attachment and a 15- residue peptide designated T2-15 (DCQDHSF-SIVIETVQ, residues numbered 395- 409) promoted VLA-4 dependent cell adhesion. The peptide was also capable of inhibiting cell adhesion to pp-vWF, suggesting that this sequence represents the cell attachment site. By affinity chromatography using peptide T2-15- Sepharose, it was found that α4β1 integrin complex from extracts of surface iodinated B16 cells specifically bound to the peptide. These results strongly suggest that pp-vWF is a novel physiological ligand for VLA-4.

AB - We have previously reported that propolypeptide of von Willebrand factor (pp-vWF) promotes melanoma cell adhesion in a β1 integrin-dependent manner. In this report, we identified the α subunit of the cell adhesion receptor for pp-vWF as α4. Human leukemia cell lines that express α4β1 integrin (very late antigen-4, VLA-4), but not cell lines which lack VLA-4, attached well to pp-vWF substrate and these adhesions were completely inhibited by anti-α4 integrin monoclonal antibody HP2/1. Adhesion of mouse melanoma expressing α4 integrin was also inhibited by anti-mouse α4 mAb PS/2. Furthermore, transfection of human α4 cDNA into α4-Chinese hamster ovary cells resulted in an acquisition of adhesive activity to pp-vWF, indicating that pp-vWF is a ligand for VLA-4 integrin. Using a recombinant fragment of pp-vWF, the cell attachment site was shown to be located within amino acid residues 376-455 of pp-vWF. A series of synthetic peptides covering this region were tested for the ability to promote cell attachment and a 15- residue peptide designated T2-15 (DCQDHSF-SIVIETVQ, residues numbered 395- 409) promoted VLA-4 dependent cell adhesion. The peptide was also capable of inhibiting cell adhesion to pp-vWF, suggesting that this sequence represents the cell attachment site. By affinity chromatography using peptide T2-15- Sepharose, it was found that α4β1 integrin complex from extracts of surface iodinated B16 cells specifically bound to the peptide. These results strongly suggest that pp-vWF is a novel physiological ligand for VLA-4.

UR - http://www.scopus.com/inward/record.url?scp=0030887708&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030887708&partnerID=8YFLogxK

U2 - 10.1074/jbc.272.13.8447

DO - 10.1074/jbc.272.13.8447

M3 - Article

C2 - 9079671

AN - SCOPUS:0030887708

VL - 272

SP - 8447

EP - 8453

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 13

ER -