Functional domains of the 67-kDa laminin receptor precursor

Vincent Castronovo, Giulia Taraboletti, Mark E. Sobel

Research output: Contribution to journalArticle

110 Citations (Scopus)

Abstract

We report the characterization of two functional domains of the metastasis-associated 67-kDa laminin receptor (67-LR). Using synthetic peptides deduced from the cDNA sequence of the 37-kDa precursor of the laminin receptor (37-LRP) as well as their corresponding affinity-purified polyclonal antibodies, we identified a unique laminin binding site as well as a membrane-associated domain of the receptor. In laminin dot blot and solid phase radioligand assays, a 20 amino acid synthetic peptide (IPCNNKGAHSV-GLMWWMLAR, amino acid residues 161-180, designated peptide G) specifically bound to laminin with high affinity (Kd = 5 × 10-8 M). Peptide G also specifically eluted the 67-LR from a laminin affinity column. Peptide G and laminin reacted with a 1:1 stoichiometry, suggesting that there is one recognition site on laminin for the peptide G domain. Immunofluorescence studies, performed on permeabilized and nonpermeabilized human A2058 melanoma cells using 10 different affinity-purified antibodies to distinct regions of the 37-LRP, identified an unusually short membrane-associated domain that was consistent with a computer predicted transmembrane domain (residues 86-101). Our data demonstrate for the first time that the 37-LRP has two functional domains consistent with the characteristics of the mature 67-LR. Furthermore, we propose peptide G as a potential inhibitor of tumor cell interactions with laminin.

Original languageEnglish
Pages (from-to)20440-20446
Number of pages7
JournalJournal of Biological Chemistry
Volume266
Issue number30
Publication statusPublished - 1991

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Laminin Receptors
Laminin
Peptides
Membranes
Amino Acids
Radioligand Assay
Antibody Affinity
Antibodies
Cell Communication
Stoichiometry
Fluorescent Antibody Technique
Tumors
Melanoma
Assays
Complementary DNA
Binding Sites
Cells
Neoplasm Metastasis

ASJC Scopus subject areas

  • Biochemistry

Cite this

Functional domains of the 67-kDa laminin receptor precursor. / Castronovo, Vincent; Taraboletti, Giulia; Sobel, Mark E.

In: Journal of Biological Chemistry, Vol. 266, No. 30, 1991, p. 20440-20446.

Research output: Contribution to journalArticle

Castronovo, V, Taraboletti, G & Sobel, ME 1991, 'Functional domains of the 67-kDa laminin receptor precursor', Journal of Biological Chemistry, vol. 266, no. 30, pp. 20440-20446.
Castronovo, Vincent ; Taraboletti, Giulia ; Sobel, Mark E. / Functional domains of the 67-kDa laminin receptor precursor. In: Journal of Biological Chemistry. 1991 ; Vol. 266, No. 30. pp. 20440-20446.
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