Effects of fibronectin-type V collagen recombinant fusion protein on cell adhesion and cell proliferation.
An expression vector pTF7520-Col-V-In, which encodes a fusion protein of the cell-binding domain of fibronectin (C277) and the insulin- and heparin-binding domain of the alpha 1 chain of human type V collagen, was constructed. E. coli transfected with this plasmid synthesized a 50-kDa fusion protein. This fusion protein, C277-V, was purified from the crude extract by a single step heparin HPLC. Similar amounts of insulin bound to purified C277-V and to the alpha 1 chain of type V collagen as judged by the binding of peroxidase-conjugated insulin. Cell-adhesive activity of C277-V was lower than that of the original fibronectin fragment C274, but similar numbers of cells adhered to both protein substrates when the culture dishes were coated with 1 mM of each protein. Insulin bound to the C277-V substratum stimulated the growth of mouse mammary tumor MTD cells in serum-free culture medium.
Hatai
M
M
Biology Division, National Cancer Center Research Institute, Tokyo, Japan.
Takahara
K
K
Hashi
H
H
Kato
I
I
Yaoi
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002448:Cell Adhesion / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D003094:Collagen / Q000235:genetics*; D005353:Fibronectins / Q000235:genetics*; D005786:Gene Expression Regulation / Q000187:drug effects; D005815:Genetic Code / Q000235:genetics*; D006735:Horseradish Peroxidase; D007328:Insulin / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D005353:Fibronectins; D007328:Insulin; D011993:Recombinant Fusion Proteins; D003094:Collagen; D006735:Horseradish Peroxidase