Peptide sequences selected by BA4, a tropoelastin-specific monoclonal antibody, are ligands for the 67-kilodalton bovine elastin receptor.

A 67-kDa cell-surface elastin/laminin receptor is expressed by fetal bovine ligamentum nuchae fibroblasts and neutrophils. Two hexapeptides, VGVAPG and PGAIPG, contained within hydrophobic domains of tropoelastin are binding sites for this receptor. Studies of recombinant tropoelastin proteins and synthetic peptides demonstrated that a monoclonal antibody, BA4, recognized peptide sequences similar to those recognized by the 67-kDa receptor. Taking advantage of this similarity, an "epitope library" containing random hexapeptides was screened with BA4. Four BA4-selected peptides (VGAMPG, VGMAPG, VGSLPG, and VGLSPG) were synthesized; studies of fibroblast and neutrophil migration support the hypothesis that these peptides are ligands of the 67-kDa receptor present on ligamentum nuchae fibroblasts and neutrophils. Two additional, physically similar tropoelastin peptides,AGAIPG and PGAVGP, were also identified as peptide ligands, and hence potential binding sites within tropoelastin, of the elastin receptor. These data suggest that the 67-kDa elastin/laminin receptor may interact with a wide range of structurally similar peptides containing amino acid substitutions involving small nonpolar and uncharged amino acids.

[1]  L. Randall,et al.  No specific recognition of leader peptide by SecB, a chaperone involved in protein export. , 1990, Science.

[2]  G. P. Smith,et al.  Antibody-selectable filamentous fd phage vectors: affinity purification of target genes. , 1988, Gene.

[3]  R. Mecham,et al.  Chemotactic activity of elastin-derived peptides. , 1980, The Journal of clinical investigation.

[4]  R. Mecham,et al.  Kinetics of receptor-mediated binding of tropoelastin to ligament fibroblasts. , 1988, The Journal of biological chemistry.

[5]  R. Mecham,et al.  Neutrophils show chemotaxis to type IV collagen and its 7S domain and contain a 67 kD type IV collagen binding protein with lectin properties. , 1989, American journal of respiratory cell and molecular biology.

[6]  R. Mecham,et al.  Characterization of biologically active domains on elastin: identification of a monoclonal antibody to a cell recognition site. , 1986, Biochemistry.

[7]  J. Sambrook,et al.  Protein folding in the cell , 1992, Nature.

[8]  R. Pierce,et al.  Mammalian tropoelastin: multiple domains of the protein define an evolutionarily divergent amino acid sequence. , 1991, Matrix.

[9]  R. Mecham,et al.  Development of immunoreagents to ciliary zonules that react with protein components of elastic fiber microfibrils and with elastin-producing cells. , 1988, Biochemical and biophysical research communications.

[10]  R. Mecham,et al.  Chemotactic responses of fibroblasts to tropoelastin and elastin-derived peptides. , 1982, The Journal of clinical investigation.

[11]  R. Mecham,et al.  Antigenicity of elastin: characterization of major antigenic determinants on purified insoluble elastin. , 1982, Biochemistry.

[12]  J. Rothman,et al.  Peptide-binding specificity of the molecular chaperone BiP , 1991, Nature.

[13]  R. Mecham.  Receptors for laminin on mammalian cells , 1991, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[14]  A. McMichael,et al.  A critical role for conserved residues in the cleft of HLA-A2 in presentation of a nonapeptide to T cells. , 1992, Science.

[15]  L. Liotta,et al.  The elastin receptor shows structural and functional similarities to the 67-kDa tumor cell laminin receptor. , 1989, The Journal of biological chemistry.

[16]  S. Barondes,et al.  The elastin receptor: a galactoside-binding protein. , 1988, Science.

[17]  P. A. Peterson,et al.  Emerging principles for the recognition of peptide antigens by MHC class I molecules. , 1992, Science.

[18]  R. Mecham,et al.  Fibroblast adhesion to recombinant tropoelastin expressed as a protein A-fusion protein. , 1991, The Biochemical journal.

[19]  J. Scott,et al.  Searching for peptide ligands with an epitope library. , 1990, Science.

[20]  R. Mecham,et al.  Val-Gly-Val-Ala-Pro-Gly, a repeating peptide in elastin, is chemotactic for fibroblasts and monocytes , 1984, The Journal of cell biology.

[21]  R. Mecham,et al.  Elastin synthesis by ligamentum nuchae fibroblasts: effects of culture conditions and extracellular matrix on elastin production , 1981, The Journal of cell biology.

[22]  L. Randall,et al.  Unity in function in the absence of consensus in sequence: role of leader peptides in export. , 1989, Science.

[23]  P. A. Peterson,et al.  Crystal structures of two viral peptides in complex with murine MHC class I H-2Kb. , 1994, Science.

[24]  R. Mecham,et al.  Elastin binds to a multifunctional 67-kilodalton peripheral membrane protein. , 1989, Biochemistry.

[25]  L. Grosso,et al.  PGAIPG, a repeated hexapeptide of bovine tropoelastin, is a ligand for the 67-kDa bovine elastin receptor. , 1993, Matrix.

[26]  J. Rothman,et al.  Peptide binding and release by proteins implicated as catalysts of protein assembly. , 1989, Science.