Pka

Taskén, Kjetil; Solberg, Rigmor; Foss, Kari Bente; Sklhegg, Bjørn S.; Hansson, V.; Jahnsen, Tore

doi:10.1016/b978-012324719-3/50005-4

Cited by 9 publications

(5 citation statements)

References 21 publications

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“…at 2.0 mM EGTA) or in the presence of 200 M CaCl 2 and 50 nM calmodulin. We also measured cAMP hydrolytic activity in the presence of 0.1 and 1.0 M cAMP, two concentrations that are likely to be physiologically relevant based on the published K m values for the cAMP-dependent protein kinase (27), in order to examine the possible effects of increases in [cAMP] that might result from increased ␤-adrenergic receptor stimulation. Such increases would be expected to increase the contribution of other cAMP phosphodiesterases whose affinity for cAMP is lower than that of PDE3.…”

Section: Resultsmentioning

confidence: 99%

Isoforms of Cyclic Nucleotide Phosphodiesterase PDE3 and Their Contribution to cAMP Hydrolytic Activity in Subcellular Fractions of Human Myocardium

Hambleton

Krall

Tikishvili

et al. 2005

Journal of Biological Chemistry

102

105

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Section: Resultsmentioning

confidence: 99%

Isoforms of Cyclic Nucleotide Phosphodiesterase PDE3 and Their Contribution to cAMP Hydrolytic Activity in Subcellular Fractions of Human Myocardium

Hambleton

Krall

Tikishvili

et al. 2005

Journal of Biological Chemistry

102

105

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“…2 Briefly, a fragment of the human RII␣ cDNA sequence was amplified by polymerase chain reaction to generate a full-length open reading frame with convenient cloning sites, subcloned into the expression vector pGEX-KG, expressed as a fusion protein with GST in the Escherichia coli strain BL21/DE3, and purified from bacterial lysates by absorption to glutathione-agarose beads as described elsewhere (21,25). Subsequently, purified fusion protein was digested with thrombin, and GST was absorbed on glutathione-agarose beads to obtain soluble RII␣ protein that contained an extra two-amino acid N-terminal extension from the linker segment/thrombin cleavage site of GST.…”

Section: Methodsmentioning

confidence: 99%

“…From these observations, we can conclude that RII␣ was phosphorylated on at least two different sites during mitosis, one most likely corresponding to the autophosphorylation site and the other localized in the N-terminal domain of the protein. Analysis of the human RII␣ sequence showed a putative phosphorylation site (P) for cyclin-directed kinases in the sequence (Pro 51 -AlaAla-Thr 54 -P-Arg-Gln 57 ) of the N-terminal domain (2). Human wild type RII␣ was an in vitro substrate for CDK1, and upon phosphorylation, changed its electrophoretic mobility from 51 to 53 kDa.…”

Section: Fig 3 In Vivo Phosphorylated Rii␣ Analysis By Immunoprecipmentioning

confidence: 99%

See 1 more Smart Citation

Mitosis-specific Phosphorylation and Subcellular Redistribution of the RIIα Regulatory Subunit of cAMP-dependent Protein Kinase

Keryer

Yassenko

Labbé

et al. 1998

Journal of Biological Chemistry

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54 . Whereas RII␣ was associated with the Golgi-centrosomal region during interphase, it was dissociated from its centrosomal localization at metaphaseanaphase transition. Furthermore, particulate RII␣ from HeLa cell extracts was solubilized following incubation with CDK1 in vitro. Our results suggest that at the onset of mitosis, CDK1 phosphorylates RII␣, and this may alter its subcellular localization.Cyclic AMP-dependent protein kinases (PKAs) 1 are present in mammalian tissues as two major isozymes, type I and type II (for reviews, see Refs 1 and 2). The inactive holoenzyme is composed of a regulatory subunit dimer that binds two catalytic subunits. Binding of cAMP to the regulatory subunits results in dissociation of the catalytic subunits, which are then catalytically active and can also be translocated to the nucleus.Four different regulatory subunits (RI␣, RI␤, RII␣, RII␤) and three different catalytic subunits (C␣, C␤, C␥) have been identified as separate gene products. The regulatory subunits RI␣ and RII␣ are present in almost all cell types, whereas the expression of RI␤ and RII␤ is tissue-specific. RII isoforms are associated with A kinase-anchoring proteins (AKAPs) (3) that target PKA type II to organelles, cytoskeleton, and membranes (3-5). Intracellular accumulation of RII has been observed in association with centrosome or the Golgi complex in several mammalian cell types (6 -8).The activities and subcellular localization of protein kinases and phosphatases are known to be regulated by phosphorylation (for a review, see Ref. 9). A major difference between RI and RII is the ability of RII to be autophosphorylated by the catalytic subunit of PKA (10). Phosphorylation sites for casein kinase II, glycogen synthase kinases 3 and 5, and prolinedirected protein kinase have been identified in vitro on bovine RII␣ as well (11-13). More recently it was shown that bovine RII␤ can also be phosphorylated at Thr 69 in vitro by CDK1, the mitotic kinase p34 cdc2 associated with cyclinB (14). The physiological role of these phosphorylations is not fully understood. In vitro autophosphorylation of PKA type II decreases the affinity between RII and the catalytic subunit, and CDK1 phosphorylation of RII␤ decreases its ability to anchor to MAP-2 (a neuronal microtubule-associated protein (14)). However, RII␤ is expressed primarily in neuro-endocrine cells and is thus present in differentiated cells that divide slowly or not at all. For these reasons, we examined the cell cycle-dependent phosphorylation of the ubiquitously expressed RII␣. In the present study, we show that in dividing cells, the regulatory subunit RII␣ demonstrates different phosphorylation patterns during the cell cycle. In mitotic-arrested cells, RII␣-labeling with 8-azido[ 32 P]cAMP, immunostaining with a specific antibody, and in situ incorporation of [ 32 P]orthophosphate in HeLa cells revealed different states of RII␣ phosphorylation compared with that observed during interphase. Inhibition of PKA activity with a selective inhibitor, H89, or...

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“…Thus, no intronic sequences were expected to dimer and two free active C subunits. In human, four interrupt the 3-nontranslated region of the human different regulatory subunits (RIa, RIb, RIIa, and gene between the positions of the primers, since exonRIIb) and three distinct catalytic subunits (Ca, Cb, and intron juctions are highly conserved in cAK genes beCg) have been identified as separate gene products by tween rodents and primates (R. Solberg, K. Taské n, molecular cloning techniques (for references, see (26)). …”

Section: Product As Probes Localized the Human Ca Gene Tomentioning

confidence: 98%

The Gene Encoding the Catalytic Subunit Cα of cAMP-Dependent Protein Kinase (Locus PRKACA) Localizes to Human Chromosome Region 19p13.1

et al. 1996

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Pka

Cited by 9 publications

References 21 publications

Isoforms of Cyclic Nucleotide Phosphodiesterase PDE3 and Their Contribution to cAMP Hydrolytic Activity in Subcellular Fractions of Human Myocardium

Isoforms of Cyclic Nucleotide Phosphodiesterase PDE3 and Their Contribution to cAMP Hydrolytic Activity in Subcellular Fractions of Human Myocardium

Mitosis-specific Phosphorylation and Subcellular Redistribution of the RIIα Regulatory Subunit of cAMP-dependent Protein Kinase

The Gene Encoding the Catalytic Subunit Cα of cAMP-Dependent Protein Kinase (Locus PRKACA) Localizes to Human Chromosome Region 19p13.1

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