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C A Price

Publications and source records attributed to C A Price.

At least 109 records · Page 6Linked to original sources
PubMed1984

Synthesis of the psbA Gene Product in Euglena: In Organello and In Vitro.

To identify the psbA gene product of Euglena gracilis, we compared products translated in organello and in vitro. The most prominently labeled membrane protein of isolated Euglena plastids migrates as a band at 28 kilodaltons. An apparent precursor appears at 30 kilodaltons under conditions which inhibit the synthesis of cytoplasmically synthesized proteins. Translation of the 14S mRNA selected by hybridization with the Sephacryl S-500-immobilized psbA gene, however, yields products of approximately 37- and 41-kilodaltons. In organello, no significant label migrates to this region of the gel. We interpret these data to indicate that the primary translation product of Euglena psbA gene is larger than that of higher plants, but the mature, processed polypeptide is smaller.

Journal Article↗
PubMed1984

Effect of cimetidine on net ion fluxes across the rat gastric mucosa during mucosal damage after gastric ischemia and after intravenous acetylsalicylic acid.

This study was performed to determine the relationship between net ion fluxes across the rat gastric mucosa and gastric mucosal damage after gastric ischemia and after intravenous administration of acetylsalicylic acid, and to determine the effect of cimetidine on these parameters. Gastric mucosal blood, albumin, and fluid loss that occurred after gastric ischemia was not associated with an increase in net H+ flux from mucosa to serosa. A significant increase in net Na+ and K+ flux from serosa to mucosa occurred. In the presence of cimetidine (150 mg/kg X h, i.v.), which reduced gastric damage, net H+ flux increased significantly, whereas there was an inhibition of Na+ and K+ fluxes. Acetylsalicylic acid-induced gastric hemorrhage preceded an increase in net H+ flux from serosa to mucosa, although net Na+ and K+ fluxes were unaffected. Cimetidine (150 mg/kg X h, i.v.) potentiated gastric hemorrhage and this was associated with an increase in net K+ flux from serosa to mucosa. These studies demonstrate that overt damage to the rat gastric mucosa can occur without changes in net ion fluxes across the gastric mucosa.

Animals↗
PubMed1983

Plastid translation in organello and in vitro during light-induced development in Euglena.

Plastids were isolated from dark-grown Euglena gracilis, from cells exposed to light for 3 h, and from normal, light-grown cells. Plastid protein synthesis was then measured either in organello, by incorporation of [35S]Met into isolated plastids, or in vitro, by programming wheat germ, reticulocyte, or Escherichia coli translation systems with RNA isolated from the plastids. Specific and total rates of protein synthesis in organello increased during light-induced development about 100-fold. In contrast, specific mRNA activity of plastid RNA increased no more than 3-fold, and the estimated total mRNA activity of plastids increased less than 10-fold. Protein synthesis in plastids appears therefore to be subject to strong regulation at a level other than that of transcription superimposed on weak transcriptional regulation. The synthesis of the large subunit of ribulose-bisphosphate carboxylase in particular appears to be under translational control. There is also qualitative evidence for regulation. Most plastid mRNAs, as indicated by translation in vitro, electrophoresis, and fluorography of their translation products, increase with light-induced development as a coordinate set. A second or proplastid set can be detected in RNAs from immature plastids, but not from mature chloroplasts. A few mRNAs appear only later in development, corresponding to a delayed set. Most translation products measured in organello also appear to belong to the coordinate set, a few to the proplastid and delayed sets, and a further few to a transient set, which appears only early in development.

Euglena gracilis↗
PubMed1983

Cytoplasmic regulation of chloroplast translation in Euglena gracilis.

A regulatory role for cytoplasmically derived proteins in chloroplast translation in organello was examined by analyzing protein synthesis in plastids isolated from cells of Euglena gracilis which had been treated with cycloheximide (CHI). Incorporation of [35S]methionine by chloroplasts from CHI-inhibited Euglena was reduced approximately 40 and 90% by exposure of the cells to the antibiotic for 2 and 4 h, respectively. The chloroplast translation products were then analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography. The synthesis of polypeptides in the soluble compartment of the plastid was substantially diminished by as little as 15 min of CHI pretreatment. No qualitative alterations of the polypeptide pattern were detected. Qualitative changes were seen in the thylakoid fraction, however. Comparison of the stainable polypeptides and fluorographs of thylakoid membranes from CHI-treated cells with those of controls showed several instances in which the more slowly migrating member of a doublet accumulated with a concomitant depletion of a more rapidly migrating component. A pair of polypeptides at 28 and 30 kDa, which we believe are the Euglena homologs of the photogene product and its precursor, respectively, are representative of this phenomenon. Additionally, thylakoids from cells pretreated with CHI sometimes synthesized novel polypeptides larger than 65 kDa. Finally, when intact chloroplasts from CHI-inhibited Euglena were incubated with a postchloroplast supernatant from normal cells, there was a partial reversion of the anomalies seen in the fluorographs. These data are interpreted to indicate the cytoplasmic origin of one or more proteins whose function is to process chloroplast translation products.

Chloroplasts↗
PubMed1982

Translation of mRNAs for subunits of chloroplast coupling factor 1 in spinach.

The chloroplast coupling factor 1 consists of five nonidentical subunits, three of which (alpha, beta, and epsilon subunits) have been shown in several laboratories to be synthesized within chloroplasts. The site of synthesis of the remaining two (gamma and delta subunits) was investigated by analyzing products directed by spinach leaf RNAs in wheat germ and reticulocyte translation systems in vitro. It was found that poly(A)(+) RNA directs the synthesis of two distinct polypeptides, one of which is immunochemically related to the gamma subunit but is 4,000 daltons larger. The other shares antigenic sites with the delta subunit but is 8,000 daltons larger. When wheat germ or reticulocyte translation systems were programmed with RNAs from purified chloroplasts, the only products related to CF(1) that we could detect were a putative precursor of beta, 2,000 daltons larger than the mature subunit, and some smaller polypeptides, which appear to be incomplete translation products of beta. From these results it appears likely that the gamma and delta subunits are synthesized in the cytoplasm as larger precursors and that beta is synthesized within the chloroplast as a precursor.

Journal Article↗
PubMed1981

A comparison of the effects of sympathomimetic agents on gastric acid secretion by the rat stomach in vivo and in vitro.

1. The action of isoprenaline on gastric acid secretion in rats with Heidenhain pouches has been compared with its action in a rat isolated stomach preparation. 2. Isoprenaline (40 micrograms kg-1 h-1) inhibited the acid secretion in response to pentagastrin (20 micrograms kg-1 h-1) in conscious rate with Heidenhain pouches. 3. This inhibition could be abolished by propranolol (2 mg kg-1) and butoxamine (8 mg kg-1) and partially reversed by practolol (8 mg kg-1). 4. Propranolol (2 mg kg-1) significantly increased the response to pentagastrin (20 micrograms kt-1 h-1) but butoxamine and practolol (both at 8 mg kg-1) and the inactive isomer (+)-propranolol (2 mg kg-1) were without any effect on the pentagastrin response in the rats with pouches. 5. In the rat isolated stomach preparation isoprenaline stimulated acid secretion over the range 10(-7) M-10(-3) M whereas phenylephrine and methoxamine were without effect. 6. Propranolol (2 X 10(-5) M) inhibited this stimulatory effect of isoprenaline in vitro but (+)-propranolol (2 X 10(-5) M), practolol and butoxamine (both at 10(-4) M) had no effect on the response. 7. Propranolol (2 X 10(-5) M) did not have any effect on the response of the isolated stomach to pentagastrin (5 X 10(-7) M) or bethanechol (1.7 X 10(-5) M). 8. Phenylephrine (2 X 10(-5) M) did not affect the in vitro responses to pentagastrin (2.17 X 10(-7) M), bethanechol (1.7 X 10(-5) M) or histamine (5.4 X 10(-5) M). 9. It is concluded that isoprenaline has a direct stimulatory effect and an indirect inhibitory effect on gastric acid secretion in the rat. Both effects involve stimulation of beta-adrenoceptors. The relative predominance of one or other of these two opposing effects may help to explain the contradictory results in the literature regarding the actions of beta-adrenoceptor agonists on gastric acid secretion.

Adrenergic alpha-Agonists↗
PubMed1981

The action of beta-adrenoceptor agonists on acid secretion by the rat isolated stomach.

1. The action of beta-adrenoceptor agonists on acid secretion by an immature rat isolated stomach preparation has been studied. 2. Isoprenaline, salbutamol, salmefamol, adrenaline and noradrenaline all stimulated acid output over a concentration range of 2 X 10(-7) M--10(-5) M. 3. These responses were antagonized by propranolol (2 X 10(-5) M), pindolol and timolol (10(-6) M). 4. The antagonism of isoprenaline and salmefamol by propranolol was consistent with competitive inhibition. 5. Selective beta-adrenoceptor antagonists (practolol, atenolol, butoxamine and ICI 118 551) caused significant inhibition of noradrenaline-stimulated secretion but not of that due to the other agonists. 6. An adult rat isolated mucosa preparation responded to adrenaline in a similar manner to the immature stomach preparation. 7. Acid secretion stimulated by beta-adrenoceptor agonists was not antagonized by atropine (10(-5) M), metiamide (10(-4) M) or prostaglandin E2 (10(-5) M). The concentrations of these three antagonists caused marked inhibition of the responses to submaximal concentrations of bethanechol, histamine and pentagastrin respectively. 8. The results are discussed in relation to the possible mechanisms of action of beta-adrenoceptor stimulation of acid secretion: it is concluded that the response is probably mediated by beta-receptors on the parietal cell.

Adrenergic Agonists↗
PubMed1980

Synthesis of thylakoid membrane proteins by chloroplasts isolated from spinach. Cytochrome b559 and P700-chlorophyll a-protein.

Intact chloroplasts, purified from spinach leaves by sedimentation in density gradients of colloidal silica, incorporate labeled amino acids into at least 16 different polypeptides of the thylakoid membranes, using light as the only source of energy. The thylakoid products of chloroplast translation were visualized by subjecting membranes purified from chloroplasts labeled with [35S]methionine to electrophoresis in high-resolution, SDS-containing acrylamide gradient slab gels and autoradiography. The apparent mol wt of the labeled products ranged from less than 10,000 to greater than 70,000. One of the labeled products is the apoprotein of the P700-chlorophyll a-protein (CPI). The CPI apoprotein is assembled into a pigment-protein complex which is electrophoretically indistinguishable from the native CPI complex. Isolated spinach chloroplasts also incorporate [3H]leucine and [35S]methionine into cytochrome b559. The radioactive label remains with the cytochrome through all stages of purification: extraction of the thylakoid membranes with Triton X-100 and urea, adsorption of impurities on DEAE cellulose, two cycles of electrophoresis in Triton-containing polyacrylamide gels and electrophoresis in SDS-containing gradient gels. Cytochrome b559 becomes labeled with both [3H]leucine and [35S]methionine and accounts for somewhat less than 1% of the total isotopic incorporation into thylakoid protein. The lipoprotein appears to be fully assembled during the time-course of our labeling experiments.

Apoproteins↗
PubMed1980

Preparation of chloroplasts from euglena highly active in protein synthesis.

Chloroplasts can be obtained by gentle lysis or mild shear of spheroplasts of vitamin B(12)-deficient Euglena gracilis and then purified by isopycnic sedimentation on gradients of Ludox AM or Percoll. The chloroplasts appear compact and highly refractile by phase contrast or Hoffmann contrast microscopy. Upon incubation with [(3)H]leucine or [(35)S]methionine, the chloroplasts incorporate the amino acids into protein at rates that are 100-fold faster than we had previously observed with Euglena and up to 8-fold faster than with chloroplasts of spinach. Euglena chloroplasts prepared by the current procedure are thus qualitatively superior to those previously available from Euglena and at least as active in protein synthesis as chloroplasts from higher plants.

Journal Article↗
PubMed1980

Partial Characterization of a Cadmium-binding Protein from the Roots of Cadmium-treated Tomato.

A Cd-binding protein has been isolated from the roots of Cd-treated tomato plants cv. Rutgers. Almost all the Cd from a high-speed supernatant fraction was recovered in a 10,000-dalton fraction from a gel filtration column coincident with 250-nanometer absorbing material. DEAE-cellulose chromatography of this 10,000-dalton material yielded one major component, which eluted at 0.34 molar NaCl, had an absorption spectrum characteristic of metallothionein, and showed absorption changes upon acidification typical of metallothionein. Although the Cd-binding protein did not behave like metallothioneins from animal sources during gel electrophoresis at pH 8.9, a single band containing Cd and staining with Coomassie brilliant blue could be detected following electrophoresis at pH 6.9. Synthesis of the Cd-binding protein appeared to be "induced" by treatment of the plants with Cd(2+).

Journal Article↗
PubMed1979

Synthesis of poly(A)-containing RNA by isolated spinach chloroplasts.

Chloroplasts were isolated from spinach leaves and the intact chloroplasts separated by centrifugation on gradients of silica sol. Chloroplasts prepared in this way were almost completely free of cytoplasmic rRNA. The purified chloroplasts were incubated with 32PO4 in the light. The nucleic acids were then extracted and the RNA was fractionated into poly(A)-lacking RNA and poly(A)-containing RNA (poly(A)-RNA) via oligo(dT)-cellulose chromatography. The poly(A)-RNA had a mean size of approximately 18--20 S as determined by polyacrylamide gel electrophoresis. The poly(A)-RNA was digested with RNase A and RNase T1, and the resulting poly(A) segments were subjected to electrophoresis on a 10% w/v polyacrylamide gel 98% v/v formamide). Radioactivity was incorporated into both poly(A)-RNA and poly(A)-lacking RNA and into the poly(A) segments themselves. The poly(A) segments were between 10 and 45 residues long and alkaline hydrolysis of poly(A) segments followed by descending paper chromatography showed that they were composed primarily of adenine residues. There was no 32PO4 incorporation into acid-insoluble material in the dark. We conclude that isolated chloroplasts are capable of synthesizing poly(A)-RNA.

Chloroplasts↗
PubMed1978

Relative Requirements for Magnesium of Protein and Chlorophyll Synthesis in Euglena gracilis.

The relationships among Mg, growth, chlorophyll synthesis, and cytoplasmic polysome content were studied in Euglena gracilis grown in different levels of the metal. At all levels of magnesium from 20 to 1,600 mumolar, both protein and chlorophyll are formed with exponential kinetics. The apparent rates of synthesis and final yields of both components are greater at higher levels of Mg, but the rate of chlorophyll synthesis always exceeds the rate of protein formation; i.e. the most severely deficient cells contain proportionally more chlorophyll than the sufficient cells. Cytoplasmic polysomes isolated from Mg-deficient Euglena are indistinguishable from those isolated from control cells. We conclude that decreased rates of protein synthesis occur prior to and possibly are causal to decreased rates of chlorophyll synthesis, but that the mechanism of this inhibition remains unclear.

Journal Article↗