Influence of transformation by Rous sarcoma virus on the amount, phosphorylation and enzyme kinetic properties of enolase.

Eigenbrodt, Erich; Fister, Peter; Rübsamen, Helga; Friis, Robert R.

doi:10.1002/j.1460-2075.1983.tb01625.x

Cited by 29 publications

(15 citation statements)

References 44 publications

(27 reference statements)

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“…It has been reported that enolases from various sources can be phosphorylated, and that phosphorylation reduces catalytic activity (5). Alternatively, phosphorylation could be a signal for proteolysis (10).…”

Section: Resultsmentioning

confidence: 99%

“…Low mol wt compounds were removed by centrifugation of samples of clarified homogenates through 2.5-mL bed volume columns of Sephadex G-25, previously equilibrated with homogenization buffer, set in conical Corex tubes. Enzyme assays, separation of the enolase isozymes, immunochemical analyses, and all other materials and methods have been previously described (1 [1][2][3][4][5][6][7][8][9][10][11][12][13]. The data for total endosperm enolase activity were analyzed using the curve-fitting capability of the Sigmaplot 3.0 graphics program from Jandel Scientific.…”

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confidence: 99%

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A Developmental Analysis of the Enolase Isozymes from Ricinus communis

Miernyk¹,

Dennis²

1992

Plant Physiol.

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Enolase activity was measured in clarified homogenates of various tissues during the life cycle of the castor oil plant (Ricinus communis L. cv Baker 296). The proportions of total activity due to the plastid and cytosolic isozymes were determined after separation by ion-exchange chromatography. The contribution of the plastid isozyme varied from more than 30% of the total at the midpoint of endosperm development to less than 1% in mature leaves and roots. During endosperm development, enolase activity increased to a peak coincident with the maximum rate of storage lipid accumulation, then decreased to nearly undetectable levels in the mature seed. Plastid enolase protein, measured using an enzyme-linked immunosorbent assay, increased in parallel with the increase in activity but decreased less rapidly and was still easily detectable in mature seeds.The developing endosperm of the COS2 has been used as a model system to study compartmentation of the glycolytic pathway in plants (reviewed in ref. 4). In this tissue, all of the glycolytic enzymes occur as distinct isozymes, present in both the cytosol and plastids. The results of surveys of other plant systems have, in essence, agreed with this model (7,13). Most research on the control ofglycolytic activity has addressed the acute, short-term regulatory properties of a few key enzymes: hexokinase, phosphofructokinase, and pyruvate kinase. There have been a few partial developmental analyses of glycolytic activity during COS maturation (3,8,17) clarified by centrifugation as previously described (11). In Ricinus, it is difficult to determine the precise date of pollination. Hence, a staging system was developed, based upon seed size, endosperm fresh and dry weights, percentage of total lipid, and seed coat morphology, similar to the system described for developing cotton seeds (14). For germinating endosperm, quiescent seeds were imbibed overnight in running tap water (day 0), then planted in trays containing vermiculite and placed in a glasshouse at 28°C and ambient light. To remove some pigments and to concentrate proteins, developing seedling tissues or organelle fractions were homogenized in 10 volumes of -20°C acetone, then centrifuged and the acetone decanted. The pellet was rehomogenized in 5 volumes of -20°C acetone and recentrifuged. The acetone powders were allowed to air dry in a fume hood and then stored desiccated at room temperature. Acetone powders were homogenized in a manner identical to that for native tissues, but in 10 volumes of buffer containing 2 mm DTT, 2 mM ascorbate, and 2% insoluble PVP. Low mol wt compounds were removed by centrifugation of samples of clarified homogenates through 2.5-mL bed volume columns of Sephadex G-25, previously equilibrated with homogenization buffer, set in conical Corex tubes. Enzyme assays, separation of the enolase isozymes, immunochemical analyses, and all other materials and methods have been previously described (1 1-13). The data for total endosperm enolase activity were analyzed using the curve-...

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

A Developmental Analysis of the Enolase Isozymes from Ricinus communis

Miernyk¹,

Dennis²

1992

Plant Physiol.

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“…Also, enolase has been shown to be a substrate for phosphorylation by epidermal growth factor-receptor kinase (Reiss et al, 1986) and protein kinase C (Nettelblad and Engstrom, 1987) in mammalian systems, by a Ser/Thr/Tyr kinase in yeast (Stern et al, 1991) and in Escherichiu coli (Dannelly et al, 1989). Furthermore, alterations in activity based on changes in phosphorylation state provide compelling evidence for the posttranslational regulation of enolase (Eigenbrodt et al, 1983;Nettelblad and Engstrom, 1987). Further investigations are needed to elucidate the phosphorylation status of enolase in M .…”

Section: Discussionmentioning

confidence: 99%

Posttranscriptional and Posttranslational Control of Enolase Expression in the Facultative Crassulacean Acid Metabolism Plant Mesembryanthemum crystallinum L

1995

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“…The kinase reactions were performed by incubating immunoprecipitated beads with 20 l of kinase buffer supplemented with 20 M ATP and 3 Ci of [␥- 32 P]ATP at 30°C for 30 min. To assess the IKK␣␤ and c-Src activities, 0.5 g of GST-IB␣ protein (aa 1-317) and 1 g of acetic acid-denatured enolase (40,41) were respectively added as the substrates. The reaction mixtures were analyzed by 12% SDS-PAGE followed by autoradiography.…”

Section: Immunoprecipitation and Protein Kinase Assaysmentioning

confidence: 99%

c-Src Mediates Thrombin-Induced NF-κB Activation and IL-8/CXCL8 Expression in Lung Epithelial Cells

Lin

Cheng

Hsu

et al. 2006

The Journal of Immunology

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In this study, we examined the regulation of NF-κB activation and IL-8/CXCL8 expression by thrombin in human lung epithelial cells (EC). Thrombin caused a concentration-dependent increase in IL-8/CXCL8 release in a human lung EC line (A549) and primary normal human bronchial EC. In A549 cells, thrombin, SFLLRN-NH2 (a protease-activated receptor 1 (PAR1) agonist peptide), and GYPGQV-NH2 (a PAR4 agonist peptide), but not TFRGAP-NH2 (a PAR3 agonist peptide), induced an increase in IL-8/CXCL8-luciferase (Luc) activity. The thrombin-induced IL-8/CXCL8 release was attenuated by d-phenylalanyl-l-prolyl-l-arginine chloromethyl ketone (a thrombin inhibitor), U73122 (a phosphoinositide-phospholipase C inhibitor), Ro-32-0432 (a protein kinsase C α (PKCα) inhibitor), an NF-κB inhibitor peptide, and Bay 117082 (an IκB phosphorylation inhibitor). Thrombin-induced increase in IL-8/CXCL8-Luc activity was inhibited by the dominant-negative mutant of c-Src and the cells transfected with the κB site mutation of the IL-8/CXCL8 construct. Thrombin caused time-dependent increases in phosphorylation of c-Src at tyrosine 416 and c-Src activity. Thrombin-elicited c-Src activity was inhibited by Ro-32-0432. Stimulation of cells with thrombin activated IκB kinase αβ (IKKαβ), IκBα phosphorylation, IκBα degradation, p50 and p65 translocation from the cytosol to the nucleus, NF-κB-specific DNA-protein complex formation, and κB-Luc activity. Pretreatment of A549 cells with Ro-32-4032 and the dominant-negative mutant of c-Src DN inhibited thrombin-induced IKKαβ activity, κB-Luc activity, and NF-κB-specific DNA-protein complex formation. Further studies revealed that thrombin induced PKCα, c-Src, and IKKαβ complex formation. These results show for the first time that thrombin, acting through PAR1 and PAR4, activates the phosphoinositide-phospholipase C/PKCα/c-Src/IKKαβ signaling pathway to induce NF-κB activation, which in turn induces IL-8/CXCL8 expression and release in human lung EC.

show abstract

Influence of transformation by Rous sarcoma virus on the amount, phosphorylation and enzyme kinetic properties of enolase.

Cited by 29 publications

References 44 publications

A Developmental Analysis of the Enolase Isozymes from Ricinus communis

A Developmental Analysis of the Enolase Isozymes from Ricinus communis

Posttranscriptional and Posttranslational Control of Enolase Expression in the Facultative Crassulacean Acid Metabolism Plant Mesembryanthemum crystallinum L

c-Src Mediates Thrombin-Induced NF-κB Activation and IL-8/CXCL8 Expression in Lung Epithelial Cells

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