Purification of Basal Ganglia Phosphoproteins

Nairn, Angus C.

Abstract

It is proposed to study the role of cyclic AMP-dependent protein phosphorylation in mediating the physiological actions of dopamine in the basal ganglia. Specific substrates for cyclic AMP-dependent protein kinase which have previously been found to be highly enriched in the basal ganglia will be purified and their biochemical properties characterized. Previous studies had identified nine such proteins, of which three termed DARPP-32, ARPP-21 and ARPP-16 have been purified and the function of one, DARPP-32, has been elucidated. The remaining five proteins will be purified and their biochemical properties, including the elucidation of their primary structure, will be determined. Antibodies to each protein will be prepared and immunological techniques developed for their quantitative determination. In addition, site-directed antibodies will be prepared which specifically recognize either the dephosphorylated or phosphorylated form of each protein. These antibodies will be used for the quantitative determination of the state of phosphorylation of each protein. Previously uncharacterized protein kinases which phosphorylate DARPP-32 and ARPP-16 will be purified and characterized. Phosphoprotein phosphatase in the basal ganglia will also be purified and characterized. The characterization of basal-ganglia specific phosphoproteins will permit the development of procedures to study the role of these proteins in certain mental and neurological illnesses.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Institute of Mental Health (NIMH)
• Type: Research Program Projects (P01)
• Project #: 5P01MH040899-07
• Application #: 3860255
• Support Year: 7
• Fiscal Year: 1991

Institution

• Name: Rockefeller University
• DUNS #: 071037113
• City: New York
• State: NY
• Country: United States
• Zip Code: 10065

Publications

Plattner, Florian; Hayashi, Kanehiro; Hernández, Adan et al. (2015) The role of ventral striatal cAMP signaling in stress-induced behaviors. Nat Neurosci 18:1094-100

Kimura, Toru; Han, Wonsun; Pagel, Philipp et al. (2011) Protein phosphatase 2A interacts with the Na,K-ATPase and modulates its trafficking by inhibition of its association with arrestin. PLoS One 6:e29269

Zhou, Mingming; Rebholz, Heike; Brocia, Christine et al. (2010) Forebrain overexpression of CK1delta leads to down-regulation of dopamine receptors and altered locomotor activity reminiscent of ADHD. Proc Natl Acad Sci U S A 107:4401-6

Bertran-Gonzalez, Jesus; Håkansson, Kerstin; Borgkvist, Anders et al. (2009) Histone H3 phosphorylation is under the opposite tonic control of dopamine D2 and adenosine A2A receptors in striatopallidal neurons. Neuropsychopharmacology 34:1710-20

Kuroiwa, Mahomi; Bateup, Helen S; Shuto, Takahide et al. (2008) Regulation of DARPP-32 phosphorylation by three distinct dopamine D1-like receptor signaling pathways in the neostriatum. J Neurochem 107:1014-26

Nishi, Akinori; Kuroiwa, Mahomi; Miller, Diane B et al. (2008) Distinct roles of PDE4 and PDE10A in the regulation of cAMP/PKA signaling in the striatum. J Neurosci 28:10460-71

Barbano, Paolo E; Spivak, Marina; Flajolet, Marc et al. (2007) A mathematical tool for exploring the dynamics of biological networks. Proc Natl Acad Sci U S A 104:19169-74

Borgkvist, Anders; Usiello, Alessandro; Greengard, Paul et al. (2007) Activation of the cAMP/PKA/DARPP-32 signaling pathway is required for morphine psychomotor stimulation but not for morphine reward. Neuropsychopharmacology 32:1995-2003

Bullock, S Andrew; Platholi, Jimcy; Gjyrezi, Ada et al. (2007) Differential regulation of protein phosphatase-1(I) by neurabin. Biochem Biophys Res Commun 358:140-4

Svenningsson, Per; Bateup, Helen; Qi, Hongshi et al. (2007) Involvement of AMPA receptor phosphorylation in antidepressant actions with special reference to tianeptine. Eur J Neurosci 26:3509-17