Purification and characterization of 3':5'-cyclic GMP-dependent protein kinase.

Gill, Gordon N.; Holdy, Kalman E.; Walton, Gordon M.; Kanstein, Cynthia B.

doi:10.1073/pnas.73.11.3918

Cited by 97 publications

(33 citation statements)

References 43 publications

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“…Ten ~1 of the extract was assayed for G-kinase in 100 ,ul of assay buffer consisting of 50 mM Tris-HCl, pH 7.5, 20 mM magnesium acetate, 0.2 mM [Y-~*P]ATP (200 cpm/ pmol), and either histone F2b (0.1 mg/ml) or 100 PM of peptide substrate selective for G-kinase, RKISASEFDRPL, initially described by Colbran et al [19]. All assays for G-kinase were performed in the presence of 0.9pM protein kinase inhibitor peptide (5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24). Assays were conducted at 30°C for 5 min and terminated by aliquoting histone or the peptide to Whatman 3MM filter paper or P81 phosphocellulose paper, respectively.…”

Section: Assay For G-kinase and Catalytic Domain Activitymentioning

confidence: 99%

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Expression of the catalytic domain of cyclic GMP‐dependent protein kinase in a baculovirus system

Boerth¹,

Lincoln²

1994

FEBS Letters

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The Type I cGMP-dependent protein kinase catalytic domain (residues 336671 from the Ia isoform) has been expressed as a cGMP independent kinase in a baculovirus system. Using peptide substrates, the protein retains similar substrate specificity as the native holoenzyme. The recombinant catalytic domain catalyzes the phosphorylation of histone, but does not display the inhibition using non-substrate histones which has been described for the holoenzyme. The catalytic domain is an active kinase in mammalian cells also since vascular smooth muscle cells transfected with the cDNA encoding the catalytic domain display altered morphology. The catalytic domain of G-kinase may be a useful tool for delineating the role of cGMP-mediated protein phosphorylation in cell systems.

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Section: Assay For G-kinase and Catalytic Domain Activitymentioning

confidence: 99%

“…The enzyme was first identified in arthropod tissues [4] and was subsequently purified from bovine lung [5,6]. There are two classes of G-kinase, a Type I which exists as a dimer of identical subunits, and a Type II which exists as a monomer.…”

Section: Introductionmentioning

confidence: 99%

Expression of the catalytic domain of cyclic GMP‐dependent protein kinase in a baculovirus system

Boerth¹,

Lincoln²

1994

FEBS Letters

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“…ref. 7) including adipose tissue (8). Thus far, however, covalent enzyme modification with changes in enzyme activity catalyzed by cGMP-dependent kinase has not been demonstrated.…”

mentioning

confidence: 99%

Activation of hormone-sensitive lipase and phosphorylase kinase by purified cyclic GMP-dependent protein kinase

Khoo

Sperry

Gill

et al. 1977

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

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Cyclic GMP-dependent protein kinase, purified to homogeneity from bovine lung, was shown to activate hormone-sensitive lipase partially purified from chicken adipose tissue. The degree of activation was the same as that effected by cyclic AMP-dependent protein kinase although higher concentrations of the cyclic GMP-dependent enzyme were required (relative activities expressed in terms of histone H2b phosphorylation units). Activation by cyclic AMP-dependent protein kinase was completely blocked by the heat-stable protein kinase inhibitor protein from skeletal muscle but activation by the cyclic GMP enzyme was not inhibited. Lipase fully activated by cyclic AMP-dependent protein kinase showed no further change in activity when treated with cyclic GMP-dependent protein kinase. Lipase activated by cyclic GMP-dependent protein kinase was reversibly deactivated by purified phosphorylase phosphatase (from bovine heart); full activity was restored by reincubation with cyclic GMP and cyclic GMPdependent protein kinase. Cholesterol esterase activity in the chicken adipose tissue fraction, previously shown to be activated along with the triglyceride lipase by cyclic AMP-dependent protein kinase, was also activated by cyclic GMP-dependent protein kinase. Crude preparations of hormone-sensitive triglyceride lipase from human or rat adipose tissue and cholesterol esterase from rat adrenal were also activated by cyclic GMP-dependent protein kinase. Purified hosphorylase kinase (rabbit skeletal muscle) was also shown to be activated by cyclic GMP-dependent protein kinase. The present results, together with those of other workers on histone phosphorylation, suggest that the substrate specificities of cyclic GMP-dependent and cyclic AMP-dependent protein kinase may be similar. This is discussed in the light of a model recently proposed with regard to the relationship between the subunit structures of the two kinases. The physiologic significance of the findings remains to be established.

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“…2A) but concurrently increased PKG1␣ dimer (Fig. 2C), the physiologically active form of the kinase (17,34,35), and augmented phosphorylation of VASP at Ser239 (Fig. 2B), which is an indicator of PKG activity, in the same samples.…”

Section: G6pd Inhibition or Knockdown Decreased Endogenous Productionmentioning

confidence: 79%

20-HETE-induced mitochondrial superoxide production and inflammatory phenotype in vascular smooth muscle is prevented by glucose-6-phosphate dehydrogenase inhibition

Lakhkar

Dhagia

Joshi

et al. 2016

American Journal of Physiology-Heart and Circulatory Physiology

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. 20-HETE-induced mitochondrial superoxide production and inflammatory phenotype in vascular smooth muscle is prevented by glucose-6-phosphate dehydrogenase inhibition. Am J Physiol Heart Circ Physiol 310: H1107-H1117, 2016. First published February 26, 2016 doi:10.1152/ajpheart.00961.2015 produced by cytochrome P-450 monooxygenases in NA-DPH-dependent manner is proinflammatory, and it contributes to the pathogenesis of systemic and pulmonary hypertension. In this study, we tested the hypothesis that inhibition of glucose-6-phosphate dehydrogenase (G6PD), a major source of NADPH in the cell, prevents 20-HETE synthesis and 20-HETE-induced proinflammatory signaling that promotes secretory phenotype of vascular smooth muscle cells. Lipidomic analysis indicated that G6PD inhibition and knockdown decreased 20-HETE levels in pulmonary arteries as well as 20-HETEinduced 1) mitochondrial superoxide production, 2) activation of mitogen-activated protein kinase 1 and 3, 3) phosphorylation of ETS domain-containing protein Elk-1 that activate transcription of tumor necrosis factor-␣ gene (Tnfa), and 4) expression of tumor necrosis factor-␣ (TNF-␣). Moreover, inhibition of G6PD increased protein kinase G1␣ activity, which, at least partially, mitigated superoxide production and Elk-1 and TNF-␣ expression. Additionally, we report here for the first time that 20-HETE repressed miR-143, which suppresses Elk-1 expression, and miR-133a, which is known to suppress synthetic/secretory phenotype of vascular smooth muscle cells. In summary, our findings indicate that 20-HETE elicited mitochondrial superoxide production and promoted secretory phenotype of vascular smooth muscle cells by activating MAPK1-Elk-1, all of which are blocked by inhibition of G6PD.20-HETE; TNF-␣; elk-1; mitogen-activated protein kinase 1 and 3; extracellular signal regulated kinase 2 and 1; protein kinase G; miRs 20-HYDROXYEICOSATETRAEONIC acid (20-HETE), the -hydroxylation metabolite of arachidonic acid (AA), is produced by cytochrome P-450 monooxygenases of the (CYP4A and CYP4F gene) families in an NADPH-dependent manner (49). 20-HETE is proinflammatory and it regulates vascular and renal function (49). It also plays a critical role in the pathogenesis of systemic hypertension, renal stenosis, and atherosclerosis (27,63,65). 20-HETE increased by hypoxia inhibits hypoxia-induced pulmonary artery constriction (69). However, its role in the hypoxia-induced inflammation of pulmonary arteries or lungs is yet unclear. NEWS & NOTEWORTHY 20-Hydroxyeicosatetraeonic acid (20-HETE) plays a criticalRecently, our laboratory also found that 20-HETE is involved in promoting prolonged hypoxia-induced pulmonary vasoconstriction and that glucose-6-phosphate dehydrogenase (G6PD), which is a major producer of NADPH in the cell, and cytochrome P-450 monooxygenase enzymes are functionally coupled in vascular smooth muscle tissue (unpublished observations). G6PD inhibition relaxes pulmonary arteries by decreasing intracellular calcium (20), prevents switching of vascular smooth ...

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Purification and characterization of 3':5'-cyclic GMP-dependent protein kinase.

Cited by 97 publications

References 43 publications

Expression of the catalytic domain of cyclic GMP‐dependent protein kinase in a baculovirus system

Expression of the catalytic domain of cyclic GMP‐dependent protein kinase in a baculovirus system

Activation of hormone-sensitive lipase and phosphorylase kinase by purified cyclic GMP-dependent protein kinase

20-HETE-induced mitochondrial superoxide production and inflammatory phenotype in vascular smooth muscle is prevented by glucose-6-phosphate dehydrogenase inhibition

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