EurekaMag
+ Translate
+ Most Popular
Investigations on the organic drift in North Swedish streams
Limitations of Using Microsoft Excel Version 2016 (MS Excel 2016) for Statistical Analysis for Medical Research
Phloem necrosis of coffee in Surinam
Anatomy of Mystus seenghala IV Nervous system
Integrated farming system for gardenland conditions of Coimbatore district - an over view
Tetrapleure (Tetrapleura tetraptera), an unknown African medicinal and spice plant
The origin and phylogenetic significance of the trochophoran larvae 2. evolutionary significance of the larvae of coelomate worms and mollusks
Comparison of rice bran and maize bran as feeds for growing and fattening pigs
Enterobacter amnigenus. An unusual human pathogen
Influence of Seriboost foliar application on leaf yield and leaf protein content in mulberry (Morus spp.), in relation to silkworm cocoon production
The identity of the lipstick mold of cultivated mushrooms agaricus bisporus
Advantages and disadvantages of bordeaux mixture and of lime-sulphur used on apples in the growing season
'Pan-sukh' disease of Rice in the Central Provinces
Geological age of the Ptilophyllum flora; a critical reassessment
Study of vitellogenesis in birds; physiological phases & role of folliculin in vitellogenesis
Evaluation of WCT coconut and Komadan coconut
Therapy for acne with saccharomyces boulardii
Evidence for Late Cretaceous N-S dextral shear in the west-central crystalline core, North Cascades, Washington
'Rajeshwari' - a high-yielding white seeded variety of sesame for Andhra Pradesh
Manufacture of Ricotta cheese from whey fortified with skim milk powder using different acidulants
Occurrence of Eutrichophilus mexicanus (Rudow, 1866) and Eutrichophilus lobatus (Ewing, 1936) (Phthiraptera: Trichodectidae) on Sphiggurus villosus (Cuvier, 1825) (Rodentia: Erethizontidae) in Rio de
Factors affecting fruitfulness in durian (Durio zibethinus Murr.). I. Flowering and pollination
Bronchial cuff pressure change caused by left-sided double-lumen endobronchial tube displacement
Some Biological Applications of Organometallic Compounds
The composition of pampas-grass (Cortaderia argentea.)

The consequences of introducing an autophosphorylation site into the type I regulatory subunit of cAMP-dependent protein kinase


The consequences of introducing an autophosphorylation site into the type I regulatory subunit of cAMP-dependent protein kinase



Journal of Biological Chemistry 264(17): 9807-9813



ISSN/ISBN: 0021-9258

PMID: 2656713

The type I and type II regulatory subunits of cAMP-dependent protein kinase can be distinguished by autophosphorylation. The type II regulatory subunits have an autophosphorylation site at a proteolytically sensitive hinge region, while the type I regulatory subunits have a pseudophosphorylation site. Only holoenzyme formed with type I regulatory subunits has a high affinity binding site for MgATP. In order to determine the functional consequences of regulatory subunit phosphorylation on interaction with the catalytic subunit, an autophosphorylation site was introduced into the type I regulatory subunit using recombinant DNA techniques. When Ala97 at the hinge region of the type I regulatory subunit was replaced with Ser, the regulatory subunit became a good substrate for the catalytic subunit. Stoichiometric phosphorylation occurred exclusively at Ser97. Radioactivity was incorporated primarily into the recombinant regulatory subunit when catalytic subunit and [gamma-32P]ATP were added to the total bacterial extract. Phosphorylation of the mutant regulatory subunit also occurred readily following polyacrylamide gel electrophoresis and electrophoretic transfer to nitrocellulose. Phosphorylation occurred as an intramolecular event in the absence of cAMP indicating that the hinge region of the regulatory subunit occupies the substrate recognition site of the catalytic subunit in the holoenzyme complex. Holoenzyme formed with both the wild type and mutant regulatory subunits was susceptible to dissociation in the presence of high salt; however, only the native holoenzyme was stabilized by MgATP. In contrast to the wild type holoenzyme, the affinity of the mutant holoenzyme for cAMP was not reduced in the presence of MgATP. Holoenzyme formation also was not facilitated by MgATP.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 018163228

Download citation: RISBibTeXText

Related references

Introducing an autophosphorylation site mutation in the RII-beta regulatory subunit of cAMP-dependent protein kinase abolishes the RII-beta-mediated regulatory function. FASEB Journal 8(7): A1328, 1994

Consequences of cAMP-binding site mutations on the structural stability of the type I regulatory subunit of cAMP-dependent protein kinase. Biochemistry 39(49): 15022-15031, 2000

Correlation of the cAMP binding domain with a site of autophosphorylation on the regulatory subunit of cAMP-dependent protein kinase II from porcine skeletal muscle. Journal of Biological Chemistry 254(18): 9000-9005, 1979

Selectivity and regulation of A-kinase anchoring proteins in the heart. The role of autophosphorylation of the type II regulatory subunit of cAMP-dependent protein kinase. Journal of Biological Chemistry 275(52): 41389-41395, 2000

The 44-kDa regulatory subunit of the Paramecium cAMP-dependent protein kinase lacks a dimerization domain and may have a unique autophosphorylation site sequence. Journal of Eukaryotic Microbiology 43(4): 347-356, 1996

Autophosphorylation mutant of type I regulatory subunit of cAMP-dependent protein kinase inhibits cell growth. FASEB Journal 9(6): A1303, 1995

Site-directed mutagenesis of the cAMP-binding sites of the recombinant type I regulatory subunit of cAMP-dependent protein kinase. Biochemical and Biophysical Research Communications 153(3): 1244-1250, 1988

Studies on the site in the regulatory subunit of type I cAMP-dependent protein kinase phosphorylated by cGMP-dependent protein kinase. Journal of Biological Chemistry 256(11): 5604-5607, 1981

Interactions between regulatory and catalytic subunits of the Candida albicans cAMP-dependent protein kinase are modulated by autophosphorylation of the regulatory subunit. Biochimica et Biophysica Acta 1542(1-3): 73-81, 2002

CAMP-dependent Protein Kinase Regulatory Subunit Type IIb. Active Site Mutations Define an Isoform-specific Network for Allosteric Signaling by cAMP. Journal of Biological Chemistry 279(8): 29-36, 2004

Type II regulatory subunit of cAMP-dependent protein kinase. Phosphorylation by casein kinase II at a site that is also phosphorylated in vivo. Journal of Biological Chemistry 257(17): 10440-10445, 1982

CAMP-dependent protein kinase regulatory subunit type IIbeta: active site mutations define an isoform-specific network for allosteric signaling by cAMP. Journal of Biological Chemistry 279(8): 7029-7036, 2004

Localization and characterization of the binding site for the regulatory subunit of type II cAMP-dependent protein kinase on MAP2. Neuron 3(5): 631-638, 1989

HIP/PAP, a C-type lectin overexpressed in hepatocellular carcinoma, binds the RIIa regulatory subunit of cAMP-dependent protein kinase and alters the cAMP-dependent protein kinase signalling. European Journal of Biochemistry 271(19): 12-20, 2004

HIP/PAP, a C-type lectin overexpressed in hepatocellular carcinoma, binds the RII alpha regulatory subunit of cAMP-dependent protein kinase and alters the cAMP-dependent protein kinase signalling. European Journal of Biochemistry 271(19): 3812-3820, 2004