Identification and characterization of an intracellular protein complex that binds fibroblast growth factor-2 in bovine brain.
The fibroblast growth factor (FGF) family is composed of polypeptides with sequence identity which signal through transmembrane tyrosine kinase receptors. We report here the purification from bovine brain microsomes of an FGF-2-binding complex composed of three proteins of apparent molecular masses 150 kDa, 79 kDa and 46 kDa. Only the 150 kDa and 79 kDa proteins bound FGF-2 in cross-linking and ligand-blotting experiments. Binding of FGF-2 to p79 is enhanced in the presence of calcium. Peptide sequences allowed the identification of p150 and the cloning of the cDNAs encoding p79 and p46. The deduced amino acid sequence of p79 reveals high similarity to those of gastrin-binding protein and mitochondrial enoyl-CoA hydratase/hydroxyacyl-CoA dehydrogenase. p46 is similar to mitochondrial ketoacyl-CoA thiolase. Stable transfection of FR3T3 rat fibroblast cells with p79 cDNA analysed by electron microscopy following immunolabelling of ultra-thin cryosections revealed a localization of p79 in the secretory pathway, mainly in the endoplasmic reticulum and the Golgi region, where it is specifically associated with the molecular chaperone calnexin. In vivo a protein similar to the Golgi protein MG-160 forms a complex with FGF-2 and p79.
Chevet
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E
UER de Science, Université Paris XII, 61 avenue du Général De Gaulle, 94010 Créteil Cédex, France.
Lemaître
G
G
Cailleret
K
K
Dahan
S
S
Bergeron
J J
JJ
Katinka
M D
MD
D000102:Acetyl-CoA C-Acyltransferase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000378:metabolism; D037281:Calnexin; D002352:Carrier Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D002417:Cattle; D003001:Cloning, Molecular; D018076:DNA, Complementary; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism*; D008861:Microsomes / Q000378:metabolism; D008969:Molecular Sequence Data; D011485:Protein Binding; D051381:Rats; D017386:Sequence Homology, Amino Acid
D002135:Calcium-Binding Proteins; D002352:Carrier Proteins; D018076:DNA, Complementary; D016222:Fibroblast Growth Factor 2; D037281:Calnexin; D000102:Acetyl-CoA C-Acyltransferase