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Sphingosine Kinase Interacting Protein is an A‐Kinase Anchoring Protein Specific for Type I cAMP‐Dependent Protein Kinase
Author(s) -
Kovanich Duangnapa,
van der Heyden Marcel A. G.,
Aye Thin Thin,
van Veen Toon A. B.,
Heck Albert J. R.,
Scholten Arjen
Publication year - 2010
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201000058
Subject(s) - protein kinase a , microbiology and biotechnology , biology , protein subunit , biochemistry , kinase , cytosol , mitogen activated protein kinase kinase , enzyme , gene
The compartmentalization of kinases and phosphatases plays an important role in the specificity of second‐messenger‐mediated signaling events. Localization of the cAMP‐dependent protein kinase is mediated by interaction of its regulatory subunit (PKA‐R) with the versatile family of A‐kinase‐anchoring proteins (AKAPs). Most AKAPs bind avidly to PKA‐RII, while some have dual specificity for both PKA‐RI and PKA‐RII; however, no mammalian PKA‐RI‐specific AKAPs have thus far been assigned. This has mainly been attributed to the observation that PKA‐RI is more cytosolic than the more heavily compartmentalized PKA‐RII. Chemical proteomics screens of the cAMP interactome in mammalian heart tissue recently identified sphingosine kinase type 1‐interacting protein (SKIP, SPHKAP) as a putative novel AKAP. Biochemical characterization now shows that SPHKAP can be considered as the first mammalian AKAP that preferentially binds to PKA‐RIα. Recombinant human SPHKAP functions as an RI‐specific AKAP that utilizes the characteristic AKAP amphipathic helix for interaction. Further chemical proteomic screening utilizing differential binding characteristics of specific cAMP resins confirms SPHKAPs endogenous specificity for PKA‐RI directly in mammalian heart and spleen tissue. Immunolocalization studies revealed that recombinant SPHKAP is expressed in the cytoplasm, where PKA‐RIα also mainly resides. Alignment of SPHKAPs' amphipathic helix with peptide models of PKA‐RI‐ or PKA‐RII‐specific anchoring domains shows that it has largely only PKA‐RIα characteristics. Being the first mammalian PKA‐RI‐specific AKAP with cytosolic localization, SPHKAP is a very promising model for studying the function of the less explored cytosolic PKA‐RI signaling nodes.