Association of bovine brain-derived growth factor receptor with protein tyrosine kinase activity

Abstract

Bovine brain-derived growth factor (BDGF) is very similar to endothelial cell growth factor and brain-derived acidic fibroblast growth factor in terms of pI (5.7) and molecular weight (approximately 17,000). BDGF has a wide spectrum of cell specificity, including bovine aorta endothelial cells and Swiss mouse 3T3 cells. BDGF stimulates the phosphorylation of a 135-kDa protein in plasma membranes of 3T3 cells. The optimal concentration for stimulation of phosphorylation is close to the Kd of 125I-BDGF binding to receptor, suggesting that the BDGF-stimulated 32P-labeled 135-kDa protein may be the BDGF receptor. The alkaline stability of this 32P-labeled 135-kDa phosphoprotein and phosphoamino acid analysis of the acid hydrolysates indicate that the phosphorylation occurs at tyrosine residues. The molecular size of BDGF receptor is estimated as approximately 135 kDa by cross-linking 125I-BDGF to its receptor in 3T3 cells, using a bifunctional reagent, ethylene glycolbis(succinimidylsuccinate). Both BDGF-stimulated phosphorylation and 125I-BDGF binding to receptor can be inhibited by protamine. These results suggest that the BDGF receptor is a 135-kDa protein which is associated with a protein tyrosine kinase activity.