Presence of basic fibroblast growth factor receptors in bovine brain membranes.
We described a protocol for purification of bovine brain membranes suitable to study the binding of iodinated basic fibroblast growth factor (FGF) to bovine brain membrane preparation. The binding of 125I basic FGF to brain membranes reached equilibrium within 30 min at 20 degrees C, was reversible, and displaced by an excess of unlabeled basic FGF. Scatchard analysis of the data revealed that two classes of binding sites could be detected with an apparent Kd of 30 pM and a capacity of 0.24 pmol/mg of membrane proteins for the high affinity binding site and Kd of 3 nM with a capacity of 51 pmol/mg of membrane proteins for the low affinity binding site. Cross-linking experiments of labeled basic FGF to brain membrane receptor yield the formation of a single major complex with an apparent molecular mass of 170 kDa which is similar to the value obtained for the high affinity binding site for basic FGF on target cells in tissue culture. Hence these data present the first biochemical evidence suggesting that membrane purified from bovine brain contain two classes of specific binding sites for basic FGF and confirm results described with cells grown in vitro.
Courty
J
J
Université Paris XII, Laboratoire de Biotechnologie des Cellules Eucaryotes, Creteil, France.
Dauchel
M C
MC
Mereau
A
A
Badet
J
J
Barritault
D
D
D000818:Animals; D001667:Binding, Competitive; D001923:Brain Chemistry; D002417:Cattle; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D015636:Magnesium Chloride; D008566:Membranes / Q000032:analysis; D011956:Receptors, Cell Surface / Q000032:analysis*; D017468:Receptors, Fibroblast Growth Factor
D011956:Receptors, Cell Surface; D017468:Receptors, Fibroblast Growth Factor; D015636:Magnesium Chloride; D008274:Magnesium