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Cardiac Scaffold Protein Binding Kinetics: A Role For Altered Signaling During Human Heart Disease
Author(s) -
Rababah Abeer,
Craft John,
Guillory Ashley,
Wijaya Cori,
Singh Sonal,
Fan Qiying,
Diaz Andrea Diaz,
Yin Xing,
McConnell Bradley
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.666.8
Subject(s) - phosphorylation , signal transduction , scaffold protein , microbiology and biotechnology , protein kinase a , plasma protein binding , receptor , biology , phosphodiesterase , binding site , chemistry , biochemistry , enzyme
Cardiac contractility is physiologically modulated primarily through beta‐adrenergic (β‐AR) receptor signaling. Beta‐adrenergic stimulation signals through phosphorylation of downstream proteins such as protein kinase A (PKA). PKA substrate phosphorylation is facilitated through its co‐localization with its signaling partner by A‐kinase anchoring proteins (AKAPs). In particular, mAKAP (muscle‐selective AKAP) localizes PKA and its target substrates such as phosphodiesterase‐4D3 (PDE4D3). From different genomic databases, we have recently identified fifteen human mAKAP coding non‐synonymous polymorphisms located within or near key protein binding sites critical to β‐AR receptors signaling. Seven of these mutants were cloned for the purpose of comparing whether those substitutions disrupt mAKAP binding to either the PKA binding domain or the PDE4D3. Using surface plasmon resonance (Biacore 2000) we demonstrate specific binding of wild type mAKAP to PDE4D3. Experiments were run in triplicate and as twofold serial dilutions to explore the kinetics of the interaction and analyzed using Scrubber2 with a 1:1 Langmuir model. Comparative analysis of the binding responses of mutations to mAKAP could provide important information about how these mutations modulate signaling.