Protein Kinases and Proteins Binding Adenosine 3': 5'-Monophosphate in Subcellular Fractions of Calf Ovaries. Effect of Trypsin and Protease Inhibitors on Protein Kinases

Talmadge, Kenneth W.; Bechtel, Elke; Eppenberger, Urs

doi:10.1111/j.1432-1033.1977.tb11754.x

Cited by 14 publications

(3 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The R proteins are subject to limited proteolysis [25,7,8,13]. Exposure to large or small concentrations of various proteolytic enzymes converted the regulatory subunit of protein kinase I1 quantitatively to the monomeric R' protein (molecular weight 37 000) with the retention of the full cyclic-AMP-binding capacity and affinity, while other properties of R protein including inhibition of C were lost.…”

Section: Discussionmentioning

confidence: 99%

Adenosine-3': 5'-Monophosphate-Binding Proteins from Bovine Kidney. Isolation by Affinity Chromatography and Limited Proteolysis of the Regulatory Subunit of Protein Kinase II

Weber

Hilz

1978

Eur J Biochem

View full text Add to dashboard Cite

Affinity chromatography on cyclic AMP columns allowed a two-step isolation of the cyclic-AMP-binding proteins from bovine kidney cytosol. An AMP-binding protein (apparent molecular weight zz 60000) and large amounts of a low affinity binding protein ('P35'; apparent subunit size z 35000) were obtained in practically pure form besides the high affinity binding proteins of the R type. Among the R proteins the dimer RZ of the regulatory subunit of protein kinase I1 (apparent subunit size z 54000) represented the bulk material. Small amounts of monomer, of higher aggregates, and of a protein 'P49' (subunit size ~4 9 0 0 0 ) presumably identical with the regulatory subunit of protein kinase I were also detected.The R protein fraction of kidney also contained a high affinity binding protein of smaller size (designated as R'; molecular weight z 37000) which appeared to be derived from protein Rz of protein kinase I1 by limited proteolysis. At all stages of purification, R protein and its aggregates could be quantitatively transformed into R' protein (or a closely related polypeptide) by several proteases including the relatively unspecific proteinase K. The degradation product exhibited unchanged cyclic-AMP-binding capacities but had largely lost the ability to inhibit the catalytic subunit C of protein kinase, to be phosphorylated by C, and to form a dimer. Preliminary experiments indicate that protein R' may be a natural component of kidney tissue.Mammalian tissues contain cyclic-AMP-dependent protein kinases which are composed of catalytic (C) and regulatory (R) subunits. Dissociation of the holoenzymes occurs by binding of cyclic AMP to the regulatory R proteins (for a recent review see [l]). Two types of protein kinases (type I and type 11) have been found in many tissues [2] which appear to possess identical catalytic subunits, but differ in their regulatory subunits (cf. [l]). The detection of additional cyclic-AMP-binding proteins in mammalian tissues ~~

show abstract

Section: Discussionmentioning

confidence: 99%

Adenosine-3': 5'-Monophosphate-Binding Proteins from Bovine Kidney. Isolation by Affinity Chromatography and Limited Proteolysis of the Regulatory Subunit of Protein Kinase II

Weber

Hilz

1978

Eur J Biochem

View full text Add to dashboard Cite

show abstract

“…A similar model for trypsin effect on bovine liver protein kinase has been proposed by Sugden . The presence of a smaller form of protein kinase that is presumably derived from a large holoenzyme has also been described in the calf ovary, where a proteolytic conversion of the larger to the smaller enzyme has been proposed (Talmadge et al. 1977).…”

Section: Discussionmentioning

confidence: 95%

Cyclic adenosine 3',5'-monophosphate dependent protein kinase of rat Leydig cells: physical characteristics of two holoenzymes and their subunits

Ej¹,

Dufau²,

Kj³

1978

Biochemistry

View full text Add to dashboard Cite

The interstitial cells of the rat testis contain two forms of cAMP-dependent protein kinase which are activated in vitro by low concentrations (10~*1 M) of human chorionic gonadotropin. The two cAMP-dependent holoenzymes of purified Leydig cells have been further characterized by cAMP binding and phosphokinase assay during ion-exchange chromatography, gel filtration, and sucrose density gradient centrifugation. Equilibrium binding studies performed with interstitial cell extracts and [3H]cAMP revealed a single order of binding sites with high affinity for cAMP (Ka = 109 M_ 1). After chromatography on DEAE-Sephadex and DEAE-cellulose, the two holoenzymes were identified by density gradient centrifugation as discrete peaks with sedimentation constants of 4.0 S and 6.2 S. From these values and the Stokes radii of 37.9 Á and 47.7 Á derived from gel filtration on Sephadex G-200, the estimated molecular weights of the holoenzymes were 59 600 and 116 400, respectively. The regulatory subunits of the protein kinase holoenzymes were also identified by ./Activation of cAMP1-dependent protein kinase by gonadotropic hormones has been recently demonstrated in collagcnase-dispersed interstitial cells of the rat testis (Podesta et al., 1976a,b). Enzyme activity was stimulated by incubation

show abstract

“…Dissociation, and thus activation, of the enzyme occurs by binding of cyclic AMP to the regulatory subunit (for a review see Nimmo & Cohen, 1977). The existence of additional cyclic AMP-binding proteins in mammalian tissues (Chambaut et al, 1971;Donovan & Oliver, 1972;D0skeland & Ueland, 1975;Sugden & Corbin, 1976;Talmadge et al, 1977) and the increased level of cyclic AMP-binding proteins without concomitant increase in protein kinase activity (Prasad et al, 1976) indicate that cyclic AMP-binding proteins other than the regulatory subunit of protein kinases may also be involved in cyclic AMP-mediated reactions. A cyclic AMPbinding protein not associated with a cyclic AMPdependent protein kinase has been purified and characterized from rabbit erythrocytes (Yuh & Tao, 1974).…”

mentioning

confidence: 99%

Cyclic AMP-binding and cyclic AMP-dependent protein kinase activities in the cytosol of differentiating bone marrow erythroblasts

Setchenska¹,

Vassileva-Popova²,

Arnstein³

1981

Biochemical Journal

View full text Add to dashboard Cite

Cytosolic cyclic AMP-binding capacity and cyclic AMP-dependent protein kinase activity have been studied in relation to differentiation and maturation of rabbit bone marrow erythroblasts. Using cells fractionated by velocity sedimentation at unit gravity, it was found that both activities decreased in dividing cells when calculated in terms of cell number but remained constant per cell volume. After the final cell division, cyclic AMP-dependent protein kinase activity did not change further, whereas cyclic AMP-binding capacity declined. There were no qualitative, but only quantitative, changes in the cyclic AMP-binding proteins that are present in the cytosol of developing erythroblasts. In the immature cells, the apparent KD for the interaction of binding proteins with cyclic AMP was 4 X 10(-8) M. The data suggest that changes in cyclic AMP-binding activity during differentiation of erythroid cells are due both to changes in the amount of binding proteins and in their affinity for cyclic AMP. Plasma membranes of erythroblasts were also able to bind cyclic AMP but only in dividing cells.

show abstract

Protein Kinases and Proteins Binding Adenosine 3': 5'-Monophosphate in Subcellular Fractions of Calf Ovaries. Effect of Trypsin and Protease Inhibitors on Protein Kinases

Cited by 14 publications

References 49 publications

Adenosine-3': 5'-Monophosphate-Binding Proteins from Bovine Kidney. Isolation by Affinity Chromatography and Limited Proteolysis of the Regulatory Subunit of Protein Kinase II

Adenosine-3': 5'-Monophosphate-Binding Proteins from Bovine Kidney. Isolation by Affinity Chromatography and Limited Proteolysis of the Regulatory Subunit of Protein Kinase II

Cyclic adenosine 3',5'-monophosphate dependent protein kinase of rat Leydig cells: physical characteristics of two holoenzymes and their subunits

Cyclic AMP-binding and cyclic AMP-dependent protein kinase activities in the cytosol of differentiating bone marrow erythroblasts

Contact Info

Product

Resources

About