title: Stimulation of cyclic adenosine 3',5'-monophosphate-dependent protein kinase with brain gangliosides.
abstract: The holoenzyme of cAMP-dependent protein kinase (cAMP-kinase) partially purified from the particulate fraction of rat brain was stimulated by gangliosides. Among various gangliosides tested, GM1 was most potent, giving Ka value of 19.5 microM. The maximal activation of the kinase was obtained with 100 microM GM1 using kemptide as substrate. Gangliosides inhibited the kinase activity of the catalytic subunit of cAMP-kinase. Of various substrates tested, the ganglioside-stimulated cAMP-kinase could phosphorylate microtubule-associated protein 2, synapsin I and myelin basic protein, but not histone H1 and casein. The molecular mechanisms of the stimulatory effect of gangliosides were investigated. The kinase activated with GM1 was inhibited by the addition of PKItide, a specific inhibitor for cAMP-kinase. However, GM1 did not dissociate the holoenzyme into the catalytic and regulatory subunits and did not interfere with the binding ability of cAMP to the holoenzyme. These results suggest that the gangliosides can directly activate cAMP-kinase in a different manner from cAMP.
authors.affiliation: Department of Pharmacology, Kumamoto University School of Medicine, Japan.
authors.forename: F
authors.initials: F
authors.lastname: Arakane
authors.forename: K
authors.initials: K
authors.lastname: Fukunaga
authors.forename: M
authors.initials: M
authors.lastname: Satake
authors.forename: K
authors.initials: K
authors.lastname: Miyazaki
authors.forename: H
authors.initials: H
authors.lastname: Okamura
authors.forename: E
authors.initials: E
authors.lastname: Miyamoto
mesh_terms: D000818:Animals; D001048:Aplysia / Q000737:chemistry; D001923:Brain Chemistry; D002384:Catalysis; D002848:Chromatography, DEAE-Cellulose; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000187:drug effects* / Q000378:metabolism; D004789:Enzyme Activation; D005732:Gangliosides / Q000494:pharmacology*; D006017:Glycolipids / Q000494:pharmacology*; D051381:Rats; D013268:Stimulation, Chemical; D013379:Substrate Specificity
chemical_list: D005732:Gangliosides; D006017:Glycolipids; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases