Hormone action often requires the mobilization of intracellular second messengers to activate protein kinases and phosphatases. Anchoring proteins provide a molecular framework to orient these enzymes towards selected substrates. Prototypic examples of these "signal-directing molecules" are A-kinase Anchoring Proteins (AKAPs) that sustain multi-protein signaling complexes of the cAMP dependent protein kinase (PKA) and other enzymes. AKAP79 is a multivalent anchoring protein that binds PKA, the calcium/phospholipid dependent kinase (PKC), and the calcium/calmodulin dependent phosphatase (PP2B). During the past funding period we have demonstrated that AKAP79 is recruited into larger signaling networks with individual substrates. This proposal assesses the role of AKAP79 in facilitating PKA and PP2B regulation of AMPA-type glutamate receptor trafficking and PKC mediated suppression of M- type potassium currents.
Aim 1 tests the premise that anchored PKA sustains the surface expression of AMPA receptors whereas PP2B triggers a pathway to signal internalization of the channel. Biochemical and electrophysiological techniques will determine if anchored PKA and PP2B differentially regulate the degradation of PSD-95, an adapter protein that couples AKAP79 to an AMPA receptor subunit.
Aim 2 is derived from preliminary data suggesting that AKAP79 anchors PKC to facilitate muscarinic suppression of M-type potassium channels. RNA inference techniques and the concomitant expression of mutant AKAP forms will determine if PKC anchoring is necessary for agonist dependent and suppression of M currents and if PP2B counteracts this process.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37GM048231-23
Application #
8443424
Study Section
Special Emphasis Panel (NSS)
Program Officer
Barski, Oleg
Project Start
1992-08-01
Project End
2015-03-31
Budget Start
2013-04-01
Budget End
2015-03-31
Support Year
23
Fiscal Year
2013
Total Cost
$280,334
Indirect Cost
$100,633
Name
University of Washington
Department
Pharmacology
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
Sonkusare, Swapnil K; Dalsgaard, Thomas; Bonev, Adrian D et al. (2014) AKAP150-dependent cooperative TRPV4 channel gating is central to endothelium-dependent vasodilation and is disrupted in hypertension. Sci Signal 7:ra66
Mercado, Jose; Baylie, Rachael; Navedo, Manuel F et al. (2014) Local control of TRPV4 channels by AKAP150-targeted PKC in arterial smooth muscle. J Gen Physiol 143:559-75
Gold, Matthew G; Fowler, Douglas M; Means, Christopher K et al. (2013) Engineering A-kinase anchoring protein (AKAP)-selective regulatory subunits of protein kinase A (PKA) through structure-based phage selection. J Biol Chem 288:17111-21
Scott, John D; Dessauer, Carmen W; Tasken, Kjetil (2013) Creating order from chaos: cellular regulation by kinase anchoring. Annu Rev Pharmacol Toxicol 53:187-210
Smith, F Donelson; Reichow, Steve L; Esseltine, Jessica L et al. (2013) Intrinsic disorder within an AKAP-protein kinase A complex guides local substrate phosphorylation. Elife 2:e01319
Hinke, Simon A; Navedo, Manuel F; Ulman, Allison et al. (2012) Anchored phosphatases modulate glucose homeostasis. EMBO J 31:3991-4004
Havekes, Robbert; Canton, David A; Park, Alan J et al. (2012) Gravin orchestrates protein kinase A and β2-adrenergic receptor signaling critical for synaptic plasticity and memory. J Neurosci 32:18137-49
Kosenko, Anastasia; Kang, Seungwoo; Smith, Ida M et al. (2012) Coordinated signal integration at the M-type potassium channel upon muscarinic stimulation. EMBO J 31:3147-56
Altier, Christophe; Dubel, Stefan J; Barrere, Christian et al. (2012) AKAP79 modulation of L-type channels involves disruption of intramolecular interactions in the CaV1.2 subunit. Channels (Austin) 6:157-65
Gold, Matthew G; Reichow, Steve L; O'Neill, Susan E et al. (2012) AKAP2 anchors PKA with aquaporin-0 to support ocular lens transparency. EMBO Mol Med 4:15-26

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