title: Evidence for the existence of cAMP-dependent protein kinase phosphorylation system associated with specific phosphoproteins in stable microtubules from rat cerebral cortex.
abstract: Cyclic AMP is a second messenger by which different extracellular signals are transduced into biological responses. Within the cell, most of the effects of cAMP are mediated through the cAMP protein kinase which appears to be localized in specific compartments of the cell near to their substrate proteins. In the present study, we have investigated the possible association of cAMP-dependent protein kinase, its substrate proteins and RII binding proteins in stable microtubules from rat cerebral cortex. The results show that in this fraction there is a cAMP binding protein of 52-54 kDa. This cAMP receptor is in the inactive holoenzyme form, since the addition of cAMP (5 microM) induces an increase in the endogenous phosphorylation of different stable microtubules polypeptides, which is completely inhibited in the presence of a specific protein kinase inhibitor (PKI 5-24 1 microM). Interestingly, overlay binding assay reveals that beside MAP2, 32P/R II is able to bind stable microtubule proteins of M(r) 150 and 75 kDa which, according to their electrophoretic mobility, can also be endogenous substrates for the enzyme. We conclude that cAMP-dependent phosphorylation system is indeed associated with stable microtubules from rat cerebral cortex.
authors.affiliation: Center for Neuropharmacology, Institute of Pharmacological Sciences, Milano, Italy.
authors.forename: J
authors.initials: J
authors.lastname: Perez
authors.forename: D
authors.initials: D
authors.lastname: Tinelli
authors.forename: C
authors.initials: C
authors.lastname: Cagnoli
authors.forename: P
authors.initials: P
authors.lastname: Pecin
authors.forename: N
authors.initials: N
authors.lastname: Brunello
authors.forename: G
authors.initials: G
authors.lastname: Racagni
mesh_terms: D000818:Animals; D002540:Cerebral Cortex / Q000201:enzymology*; D008297:Male; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013379:Substrate Specificity
chemical_list: D008869:Microtubule-Associated Proteins; D010750:Phosphoproteins; D011494:Protein Kinases