PKAcProtein Kinase A Catalytic Subunit
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While there was little basal association between spinophilin and NFM in a heterologous HEK293 cell line, upon overexpression of the catalytic subunit of PKA (PKAc), there was a robust and specific association between spinophilin and NF-M (Figure 5(a)).
Using a heterologous cell model, we transfected spinophilin, NF-M, and PKAc in the absence or presence of CDK5 and its activator p35.
These studies were performed in the absence of overexpressed PKAc, suggesting that even though the association between spinophilin and NF-M is somewhat transient under these conditions, spinophilin can functionally modulate NF-M phosphorylation even in the absence of PKAc.
When both spinophilin and NF-M were overexpressed in HEK293 cells in the presence of PKAc, there was a robust and specific interaction between the two proteins.
Abbreviations 6-OHDA: 6-Hydroxydopamine AUC: Area under the curve CDK5: Cyclin-dependent protein kinase 5 D1R: Dopamine D1 receptor D2R: Dopamine D2 receptor DA: Dopamine HEK293: Human embryonic kidney 293FT cells MSN: Medium spiny neuron NF-M: Neurofilament medium PD: Parkinson disease PKA: Protein kinase A PKAc: PKA catalytic subunit PP1: Protein phosphatase 1 PSD: Postsynaptic density XIC: Extracted ion chromatogram.
The transcriptional activity of NF-[kappa]B is regulated by the I[kappa]B-associated PKAc subunit through a cyclic AMP-independent mechanism.