Fibroblast adhesion to recombinant tropoelastin expressed as a protein A-fusion protein.
A bovine tropoelastin cDNA encoding exons 15-36 that includes the elastin-receptor binding site was expressed in Escherichia coli as a fusion protein with Protein A from Staphylococcus aureus. After isolation of the fusion protein by affinity chromatography on Ig-Sepharose, the tropoelastin domain was separated from plasmid-pR1T2T-encoded Protein A (Protein A') by CNBr cleavage. Cell-adhesion assays demonstrated specific adhesion to the recombinant tropoelastin. Furthermore, the data indicate that interactions involving the bovine elastin receptor mediate nuchalligament fibroblast adhesion to the recombinant protein. In agreement with earlier studies of fibroblast chemotaxis to bovine tropoelastin, nuchal-ligament fibroblast adhesion demonstrated developmental regulation of the elastin receptor.
Grosso
L E
LE
Department of Pathology, Jewish Hospital, Washington University Medical Center, St. Louis, MO.
Parks
W C
WC
Wu
L J
LJ
Mecham
R P
RP
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002448:Cell Adhesion; D005347:Fibroblasts / Q000502:physiology; D015723:Gene Library; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification; D015183:Restriction Mapping; D013205:Staphylococcal Protein A / Q000235:genetics / Q000302:isolation & purification; D014333:Tropoelastin / Q000235:genetics* / Q000302:isolation & purification
D015345:Oligonucleotide Probes; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; D014333:Tropoelastin