8UKO

cAMP-dependent protein kinase A catalytic domain in complex with voltage gated calcium channel peptide ternary complex 2

Summary for 8UKO

• Entry DOI: 10.2210/pdb8uko/pdb
• Descriptor: cAMP-dependent protein kinase catalytic subunit alpha, ARG-GLY-PHE-LEU-ARG-SER-ALA-SER-LEU-GLY-ARG-ARG-ALA-SER-PHE-HIS-LEU, MAGNESIUM ION, ... (5 entities in total)
• Functional Keywords: protein kinase, complex, ion channel, voltage gated calcium channel, cardiac channel, stress signaling, transferase
• Biological source: Mus musculus (house mouse); More
• Total number of polymer chains: 2
• Total formula weight: 42089.25

Authors

Haji-Ghassemi, O.,Van Petegem, F. (deposition date: 2023-10-14, release date: 2024-12-11, Last modification date: 2025-01-01)

Primary citation

Yoo, R.,Haji-Ghassemi, O.,Bader, M.,Xu, J.,McFarlane, C.,Van Petegem, F.
Crystallographic, kinetic, and calorimetric investigation of PKA interactions with L-type calcium channels and Rad GTPase.
J.Biol.Chem., 301:108039-108039, 2024

PubMed Abstract: β-adrenergic signalling activates cAMP-dependent protein kinase (PKA), which regulates the activity of L-type voltage-gated calcium channels such as Ca1.2. Several PKA target sites in the C-terminal tail of Ca1.2 have been identified, and their phosphorylation has been suggested to increase currents in specific tissues or heterologous expression systems. However, augmentation of Ca1.2 currents in the heart is instead mediated by phosphorylation of Rad, a small GTPase that can inhibit Ca1.2. It is unclear how each of the proposed target sites in Ca1.2 and Rad rank towards their recognition by PKA, which could reveal a preferential phosphorylation. Here we used quantitative assays on three Ca1.2 and four Rad sites. Isothermal titration calorimetry (ITC) and enzyme kinetics show that there are two Tiers of targets, with Ca1.2 residue Ser1981 and Rad residues Ser25 and Ser272 forming Tier 1 substrates for PKA. These share a common feature with two Arginine residues at specific positions that can anchor the peptide into the substrate binding cleft of PKA. In contrast, PKA shows minimal activity for the other, Tier 2 substrates, characterized by low k values and undetectable binding via ITC. The existence of two Tiers suggests that PKA regulation of the Ca1.2 complex may occur in a graded fashion. We report crystal structures of the PKA catalytic subunit with and without a Ca1.2 and test the importance of several anchoring residues via mutagenesis. Different target sites utilize different anchors, highlighting the plasticity of PKAc to recognize substrates.

PubMed: 39615689
DOI: 10.1016/j.jbc.2024.108039

Experimental method

X-RAY DIFFRACTION (2.89 Å)