Designing isoform-specific peptide disruptors of protein kinase A localization

Burns-Hamuro, Lora L.; Ma, Yuliang; Kammerer, Stefan; Reineke, Ulrich; Self, Chris N.; Cook, Charles H.; Olson, Gary L.; Cantor, Charles R.; Braun, Andreas; Taylor, Susan S.

doi:10.1073/pnas.2628038100

Cited by 117 publications

(133 citation statements)

References 50 publications

Supporting

Mentioning

123

Contrasting

Order By: Relevance

“…This mutation increases RI binding affinity 3-fold but has no effect on the RII͞D-AKAP-2 interaction (54). In an accompanying paper, the same authors used peptide array technologies to generate a high-affinity binding peptide with a 100-fold preference for RI␣ (55). Thus, peptide antagonists are now available to test the hypothesis that isoformselective kinase anchoring occurs in a cellular context.…”

Section: Discussionmentioning

confidence: 99%

Bioinformatic design of A-kinase anchoring protein- in silico : A potent and selective peptide antagonist of type II protein kinase A anchoring

Alto

Soderling

Hoshi

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

152

144

View full text Add to dashboard Cite

Compartmentalization of the cAMP-dependent protein kinase (PKA) is coordinated through association with A-kinase anchoring proteins (AKAPs). A defining characteristic of most AKAPs is a 14-to 18-aa sequence that binds to the regulatory subunits (RI or RII) of the kinase. Cellular delivery of peptides to these regions disrupts PKA anchoring and has been used to delineate a physiological role for AKAPs in the facilitation of certain cAMP-responsive events. Here, we describe a bioinformatic approach that yields an RIIselective peptide, called AKAP-in silico (AKAP-IS), that binds RII with a Kd of 0.4 nM and binds RI with a Kd of 277 nM. AKAP-IS associates with the type II PKA holoenzyme inside cells and displaces the kinase from natural anchoring sites. Electrophysiological recordings indicate that perfusion of AKAP-IS evokes a more rapid and complete attenuation of ␣-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor currents than previously described anchoring inhibitor peptides. Thus, computerbased and peptide array screening approaches have generated a reagent that binds PKA with higher affinity than previously described AKAPs.

show abstract

Section: Discussionmentioning

confidence: 99%

Bioinformatic design of A-kinase anchoring protein- in silico : A potent and selective peptide antagonist of type II protein kinase A anchoring

Alto

Soderling

Hoshi

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

152

144

View full text Add to dashboard Cite

show abstract

“…Thus, RIAD exhibits a 1760-fold preference for RI␣ over RII␣. This suggests that RIAD is 20-fold more selective for RI␣ than PV-38, a peptide derived from the dual specificity anchoring protein D-AKAP1 (21). RIAD bound with similar affinity to human and bovine RI␣ as assessed by the solid phase overlay assay (data not shown).…”

Section: Development Of An Optimal Ri␣ Binding Consensus Sequence-mentioning

confidence: 95%

“…The prototypic inhibitor that has been used to disrupt PKA localization is the peptide known as Ht31 (19), which encompasses the RII binding domain of human thyroid AKAP (AKAP-Lbc) (20). Recently, the dual specificity anchoring protein D-AKAP2 has been used as a template to design peptides that preferentially interact with RI␣ in vitro (21). Here, we have conducted a bioinformatics analysis on a set of dual-affinity AKAP binding sequences to develop a peptide that binds RI␣ with higher affinity and specificity than any naturally occurring AKAP or other RI␣-binding peptide reported.…”

mentioning

confidence: 99%

Delineation of Type I Protein Kinase A-selective Signaling Events Using an RI Anchoring Disruptor

Carlson

Lygren

Berge

et al. 2006

Journal of Biological Chemistry

139

155

View full text Add to dashboard Cite

Control of specificity in cAMP signaling is achieved by A-kinase anchoring proteins (AKAPs), which assemble cAMP effectors such as protein kinase A (PKA) into multiprotein signaling complexes in the cell. AKAPs tether the PKA holoenzymes at subcellular locations to favor the phosphorylation of selected substrates. PKA anchoring is mediated by an amphipathic helix of 14 -18 residues on each AKAP that binds to the R subunit dimer of the PKA holoenzymes. Using a combination of bioinformatics and peptide array screening, we have developed a high affinity-binding peptide called RIAD (RI anchoring disruptor) with >1000-fold selectivity for type I PKA over type II PKA. Cell-soluble RIAD selectively uncouples cAMP-mediated inhibition of T cell function and inhibits progesterone synthesis at the mitochondria in steroid-producing cells. This study suggests that these processes are controlled by the type I PKA holoenzyme and that RIAD can be used as a tool to define anchored type I PKA signaling events.The cAMP signaling pathway synchronizes a variety of physiological responses including cell proliferation and differentiation, microtubule dynamics, reproductive function, modulation of immune responses, and steroidogenesis (1-3). Many of these responses require activation of the cAMP-dependent protein kinase (PKA).3 The dormant PKA holoenzyme is a heterotetramer composed of two catalytic (C) subunits held in an inactive conformation by a regulatory (R) subunit dimer. Upon activation with cAMP, the C subunits are released from their interaction with the R subunit dimer and are free to phosphorylate a plethora of target substrates. Individual cells express a range of PKA isozymes differing in R (RI␣, RI␤, RII␣, RII␤) and C (C␣, C␤, C␥) subunit composition. The type I and type II PKA isozymes, which are classified on the basis of their R subunit dimer, possess slightly different physical and biological properties including a differential sensitivity to cAMP.Spatiotemporal regulation of PKA phosphorylation events is facilitated by A-kinase anchoring proteins (AKAPs). This family of structurally diverse but functionally related proteins act as molecular scaffolds to cluster PKA with specific substrates and signal termination enzymes such as phosphatases (4, 5) and cAMP-specific phosphodiesterases (6 -9). The number of AKAPs has been estimated to more than 75, of which ϳ50 have been identified to date (3, 10). Although the majority of AKAPs described to date bind type II PKA via the RII regulatory subunit, several dual function AKAPs are capable of interacting with both the type I and the type II PKA holoenzyme (11-15). There are also examples of anchoring proteins such as AKAPce (16), PAP7 (17), and merlin (18) that selectively interact with the RI subunit of PKA.The molecular basis for PKA anchoring is the interaction of an amphipathic helical motif of 14 -18 residues on the AKAP with a hydrophobic groove in the N-terminal dimerization domain of the R subunit. High affinity peptides mimicking the amphipathic helix bind to the R...

show abstract

“…4], suggests that AKAP10 serves a critical function in diverse species (11). The human gene is polymorphic in the PKA-binding domain at position 646, resulting in protein isoforms with different affinities for PKA that correlate with different cellular localizations (11)(12)(13). The predominant isoform, 646I, has a lower affinity for PKA than the minor isoform 646V.…”

mentioning

confidence: 99%

Gene-trapped mouse embryonic stem cell-derived cardiac myocytes and human genetics implicate AKAP10 in heart rhythm regulation

Tingley

Pawlikowska

Zaroff³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Sudden cardiac death due to abnormal heart rhythm kills 400,000 -460,000 Americans each year. To identify genes that regulate heart rhythm, we are developing a screen that uses mouse embryonic stem cells (mESCs) with gene disruptions that can be differentiated into cardiac cells for phenotyping. Here, we show that the heterozygous disruption of the Akap10 (D-AKAP2) gene that disrupts the final 51 aa increases the contractile response of cultured cardiac cells to cholinergic signals. In both heterozygous and homozygous mutant mice derived from these mESCs, the same Akap10 disruption increases the cardiac response to cholinergic signals, suggesting a dominant interfering effect of the Akap10 mutant allele. The mutant mice have cardiac arrhythmias and die prematurely. We also found that a common variant of AKAP10 in humans (646V, 40% of alleles) was associated with increased basal heart rate and decreased heart rate variability (markers of low cholinergic/vagus nerve sensitivity). These markers predict an increased risk of sudden cardiac death. Although the molecular mechanism remains unknown, our findings in mutant mESCs, mice, and a common human AKAP10 SNP all suggest a role for AKAP10 in heart rhythm control. Our stem cell-based screen may provide a means of identifying other genes that control heart rhythm.G protein ͉ receptor ͉ signaling

show abstract

Designing isoform-specific peptide disruptors of protein kinase A localization

Cited by 117 publications

References 50 publications

Bioinformatic design of A-kinase anchoring protein- in silico : A potent and selective peptide antagonist of type II protein kinase A anchoring

Bioinformatic design of A-kinase anchoring protein- in silico : A potent and selective peptide antagonist of type II protein kinase A anchoring

Delineation of Type I Protein Kinase A-selective Signaling Events Using an RI Anchoring Disruptor

Gene-trapped mouse embryonic stem cell-derived cardiac myocytes and human genetics implicate AKAP10 in heart rhythm regulation

Contact Info

Product

Resources

About