title: Neuronal protein tyrosine kinases associated with synaptosomal glycoproteins.
abstract: Protein tyrosine kinase (PTK) activity associated with synaptosomal membrane glycoprotein (SMGP) fractions of rat brain was examined. The synthetic substrate poly(Glu4-Tyr) was phosphorylated by SMGP in the presence of Mg2+ and Mn2+, whose stimulatory effects were additive. In contrast, endogenous tyrosine phosphorylation in SMGPs was strictly dependent on Mn2+. Anti-phosphotyrosine antibodies (PY20) immunoprecipitated two polypeptides in SMGPs of Mr 170K and 60K. Upon preincubation with IGF-I, 97/90K polypeptides were phosphorylated, corresponding to the IGF-I receptor beta-subunits, and were immunoprecipitated with both PY20 and anti-IGF-I-receptor antibodies. Immunoblot analysis using anti-src antibody revealed that there was src protein associated with the glycoprotein fractions of solubilized synaptosomal membranes. Additional experiments revealed that the 60K tyrosine-phosphorylated polypeptide present in the PY20 precipitates was indeed pp60c-src. This was confirmed by subjecting the PY20 immunoprecipitates to immunoblotting using anti-src antibodies. In addition, src protein was directly immunoprecipitated by anti-src antibodies from the SMGP preparations. Hence, IGF-I receptors and glycoprotein-associated PTKs including pp60c-src may play an important role in synaptic transmembrane signalling, plasticity, and neuronal survival.
authors.affiliation: Department of Cell Biology, Burroughs Wellcome Co., Research Triangle Park, North Carolina 27709.
authors.forename: S H
authors.initials: SH
authors.lastname: Hanissian
authors.forename: N
authors.initials: N
authors.lastname: Sahyoun
mesh_terms: D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D008562:Membrane Glycoproteins / Q000378:metabolism*; D009474:Neurons / Q000201:enzymology*; D010766:Phosphorylation; D011233:Precipitin Tests; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D051381:Rats; D013570:Synaptic Membranes / Q000201:enzymology / Q000378:metabolism; D013574:Synaptosomes / Q000201:enzymology*
chemical_list: D008562:Membrane Glycoproteins; D011505:Protein-Tyrosine Kinases