Article (Scientific journals)
Functional domains of the 67-kDa laminin receptor precursor.
Castronovo, Vincenzo; Taraboletti, G.; Sobel, M. E.
1991In Journal of Biological Chemistry, 266 (30), p. 20440-6
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Keywords :
Amino Acid Sequence; Animals; Blotting, Western; Chromatography, Affinity; DNA/genetics; Mice; Molecular Sequence Data; Radioligand Assay; Receptors, Immunologic/genetics/metabolism; Receptors, Laminin
Abstract :
[en] 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 (IPCNNKGAHSVGLMWWMLAR, amino acid residues 161-180, designated peptide G) specifically bound to laminin with high affinity (Kd = 5 x 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.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Castronovo, Vincenzo ;  Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Biologie générale et cellulaire - GIGA-R : Labo de recherche sur les métastases
Taraboletti, G.
Sobel, M. E.
Language :
English
Title :
Functional domains of the 67-kDa laminin receptor precursor.
Publication date :
1991
Journal title :
Journal of Biological Chemistry
ISSN :
0021-9258
eISSN :
1083-351X
Publisher :
American Society for Biochemistry and Molecular Biology, Baltimore, United States - Maryland
Volume :
266
Issue :
30
Pages :
20440-6
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 25 May 2010

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