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A Oaks

Publications and source records attributed to A Oaks.

At least 37 records · Page 2Linked to original sources
PubMed1983

Synthesis and degradation of barley nitrate reductase.

Nitrate and light are known to modulate barley (Hordeum vulgare L.) nitrate reductase activity. The objective of this investigation was to determine whether barley nitrate reductase is regulated by enzyme synthesis and degradation or by an activation-inactivation mechanism. Barley seedling nitrate reductase protein (cross-reacting material) was determined by rocket immunoelectrophoresis and a qualitative immunochemical technique (western blot) during the induction and decay of nitrate reductase activity. Nitrate reductase cross-reacting material was not detected in root or shoot extracts from seedlings grown without nitrate. Low levels of nitrate reductase activity and cross-reacting material were observed in leaf extracts from plants grown on nitrate in the dark. Upon nitrate induction or transfer of nitrate-grown etiolated plants to the light, increases in nitrate reductase activity were positively correlated with increases in immunological cross-reactivity. Root and shoot nitrate reductase activity and cross-reacting material decreased when nitrate-induced seedlings were transferred to a nitrate-free nutrient solution or from light to darkness. These results indicate that barley nitrate reductase levels are regulated by de novo synthesis and protein degradation.

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PubMed1980

cis-4-Cyclohexene-1,2-dicarboximide: Inhibitor of phytochrome-promoted seed germination.

cis-4-Cyclohexene-1,2-dicarboximide (CHDC) inhibits the germination of light-requiring seeds in both light and darkness but has no effect upon the germination of non-light-requiring seeds. In lettuce seeds, CHDC inhibits the action of far-red-absorbing form of phytochrome in breaking dormancy. This inhibition can be overcome by benzyladenine and red light together, but not by a combination of red light and gibberellic acid. Gibberellic acid-induced germination of lettuce seeds in darkness is inhibited also by CHDC. Embryos isolated from dark-imbibed lettuce seeds germinate on the inhibitor. CHDC was thought to be an "analogue" of cycloheximide, but it does not inhibit protein synthesis in lettuce seeds. Our results lead us to conclude that CHDC inhibits germination of seeds that require red light to break dormancy and interferes with some aspect of metabolism that is stimulated by far-red-absorbing form of phytochrome.

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PubMed1980

Characteristics of Nitrate Reductase-inactivating Proteins Obtained from Corn Roots and Rice Cell Cultures.

Nitrate reductase (NR)-inactivating proteins from corn roots (Wf-9 x 38-11) and rice cell suspension cultures were tested against a partially purified NR obtained from corn leaves (W64A x W182E). The corn protein was purified 921-fold and the rice protein, 1,660-fold using standard purification procedures. Approximate molecular weight values were 75,000 for the corn protein, and 150,000 for the rice protein as determined by Sephadex G-100 gel filtration. The Sephadex-treated proteins were characterized by electrophoresis on polyacrylamide gels. With a running pH of 9.4 the corn protein remained at the origin whereas the rice protein migrated with an R(F) value of 0.49. With a running pH of 4.0 the corn protein migrated with an R(F) value of 0.25. With the corn protein the activities of NR inactivation and hydrolysis of azocasein were detected in the same protein band. The rice protein, however, had no associated protease activity. From sodium dodecyl sulfate gel electrophoresis, there was one major protein band with an estimated molecular weight of 66,000 in corn protein. In rice protein four bands were observed with estimated molecular weights of 73,000, 66,000, 62,500, and 58,500, respectively.Both inactivators had an inhibitory effect on NADH-NR and NO(3) (-) induced NADH-cytochrome c reductase activities but they had less influence on the activities of FMNH(2)-NR and reduced methylviologen-NR. Inactivation of rice cell NR by rice inactivator was reversed by addition of NADH. Inactivation of corn leaf NR by rice inactivator was inhibited by the simultaneous addition of NADH, but rice inactivator-inactivated corn leaf NR could not be reactivated by NADH.

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PubMed1980

Action of Corn and Rice-inactivating Proteins on a Purified Nitrate Reductase from Chlorella vulgaris.

When nitrate reductase (NR) purified from Chlorella was incubated with NR-inactivating proteins purified from corn roots and rice cell suspension cultures or with trypsin there was a loss in NADH-NR and NADH cytochrome c reductase (NADH-CR) activities with time whereas the reduced methylviologen NR (MV-NR) remained active. When NADH-NR and NADH-CR activities were inactivated completely by the incubation with corn protein, the major protein band obtained by polyacrylamide gel electrophoresis shifted from an R(F) value of 0.12 to an R(F) of 0.25 and reduced MV-NR activity moved to the new position on the gel. When NADH-NR and NADH-CR activities were partially inactivated by the corn protein, NADH-NR activity was detected in an intermediate position (R(F) value of 0.18). Incubation with trypsin also caused a change in the NR protein migration pattern (R(F) value of 0.20). This protein band also had reduced MV-NR activity. Thus, the corn inactivator degrades NR in a fashion similar to but not identical with trypsin. The incubation of NR with rice inactivating protein resulted in a loss of NADH-NR but had no effect on the migration of NR protein or on the reduced MV-NR activity or mobility suggesting that the rice protein binds to Chlorella NR.

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PubMed1980

Activation of nitrate reductase by extracts from corn scutella.

NADH-nitrate reductase (NR) from the primary leaves and root tips of corn seedlings (var. W64A x W182E) were activated by extracts from corn scutella. The activator extracted in potassium phosphate buffer (pH 7.5) or 80% (v/v) ethanol and fractionated by Dowex 1 (acetate) and Dowex 50 (H(+)) resins was recovered in the cationic fraction. The activator was not detected in extracts from shoots, roots, or endosperm of the seedlings. It activated the nitrate-induced cytochrome c reductase of NR complex but had slight inhibitory effects on the activities of FMNH(2)-NR and reduced methylviologen-NR. In addition the activator inhibited the activities of purified NR-inactivating proteins from corn roots (var. Wf9 x 38-11) and rice cell cultures.

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PubMed1979

A comparison of glutamate synthase obtained from maize endosperms and roots.

Glutamate synthase (EC 2.6.1.53) has been examined in developing endosperms and roots of maize. KCl is required for maximum activity in each tissue. The effect with KCl is seen with buffer strength of 25 to 100 millimolar in the assay. The optimum concentration for the enzyme from endosperm is 20 millimolar and for the enzyme from root tissue the saturating concentration is about 20 millimolar. In root material the enzyme is labile but activity can be restored if KCl is added to the assay. Divalent cations such as Mg(2+) or Mn(2+) also activate the enzyme to some extent.In each case NADH or NADPH can serve as reductant. The reaction is insensitive to alpha-aminooxyacetate, but is inhibited by glutamate, the glutamate analogs methionine sulfoximine and methionine sulfone, and by the glutamine analogs azaserine and albizziin.

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PubMed1978

Effect of l-Canavanine on Nitrate Reductase in Corn Roots.

l-Canavanine inhibits the appearance of nitrate reductase (NADH-nitrate oxidoreductase, EC 1.6.6.1) in both root tips and mature root sections of corn (Zea mays L.). Ten-fold more canavanine was required to cause a 50% reduction in the level of nitrate reductase activity (NRA) in root tips than in mature root sections. For example with one particular batch of seeds 500 mum canavanine was effective in root tips whereas only 50 mum was required in mature root sections. In root tips arginine (1 mm) completely reversed the effect of 1 mm canavanine. In mature root sections higher concentrations of arginine (approximately 5 mm) were required for a complete reversal of the canavanine effect. Additions of canavanine to roots after a period of 3 hours with 5 mm KNO(3) resulted in a loss of NRA. NO(3) (-) protected nitrate reductase from this inactivation in both root tip and mature root sections.

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PubMed1977

Ammonium and amino acids as regulators of nitrate reductase in corn roots.

When amino acids or ammonia are added to plant systems, the effects on the development of nitrate-dependent nitrate reductase activity are variable. In addition, amino acids added singly or as casein hydrolysate may not support a normal growth. A physiologically correct mixture of amino acids, one similar in composition to amino acids released by the endosperm, has been shown to support normal growth and protein synthesis in corn (Zea mays) embryos. In this investigation, we have used the mixture of corn amino acids to determine whether amino acids have an effect on the appearance or disappearance of nitrate reductase activity. The results show that these amino acids partially inhibit the induction of nitrate reductase in corn roots. The effect is more pronounced in mature root than in root tip sections. When glutamine and asparagine are included along with the "corn amino acid mixture," the inhibition is more severe. Amino acids or amino acid analogues added singly to the induction medium have a similar effect: i.e. when the induction of nitrate reductase is inhibited in the root tips (lysine, canavanine, azaserine, azetidine-2-carboxylic acid, dl-4-azaleucine, asparagine, and glutamine), that inhibition is more severe in mature root sections. Arginine enhanced the recovery of nitrate reductase in root tips but inhibited it in mature root sections. The effect of the amino acids is apparently on some phase of the induction processes (i.e. the uptake or distribution of nitrate or a direct effect on the synthesis of the enzyme) and not on the turnover of the enzyme.

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PubMed1977

Asparagine synthetase in corn roots.

The level of asparagine synthetase is low in 10-mm root tips from corn seedings (Zea mays W64 x W182F) but relatively high in mature root sections taken 20 to 35 mm from the tip. When root tips are excised there is a marked increase in asparagine synthetase over a 5-hour period. In mature root sections, on the other hand, the asparagine synthetase activity declines over the same 5-hour period. The increase in the root tip is sensitive to cordycepin, 6-methylpurine, and cycloheximide, which indicates that both RNA and protein synthesis are involved in the formation of asparagine synthetase in the root tip sections. The glutamine analogue azaserine also inhibits formation of the enzyme in root tips, as does glucose. The increase in the root tip is not sensitive to asparagine. Additions of glucose or asparagine have no effect on enzyme activity in extracts. When cycloheximide, azaserine, or glucose is added to the mature root sections there is no effect on recovered enzyme activity.

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PubMed1976

Comparative studies on the induction and inactivation of nitrate reductase in corn roots and leaves.

A comparison of induction and inactivation of nitrate reductase and two of its component activities, namely FMNH(2)-nitrate reductase and NO(3) (-)-induced NADH-cytochrome c reductase, was made in roots and leaves of corn (Zea mays L. var. W64A x 182E). The three activities were induced in parallel in both tissues when NO(3) (-) was supplied. WO(4) (=) suppressed the induction of NADH- and FMNH(2)-nitrate reductase activities in root tips and leaves. The NO(3) (-)-induced NADH-cytochrome c reductase activity showed a normal increase in roots treated with WO(4) (=). In leaves, on the other hand, there was a marked superinduction of the NO(3) (-)-induced NADH-cytochrome c reductase in the presence of WO(4) (=).The half-life values of NADH-nitrate reductase and FMNH(2)-nitrate reductase measured by removing NO(3) (-) and adding WO(4) (=) to the medium, were 4 hours in root tips and 6 hours in excised leaves. Addition of NO(3) (-) in the induction medium together with WO(4) (=) gave partial protection of NADH-nitrate reductase and FMNH(2)-nitrate reductase activities in both root tips and leaves with a t(0.5) of 6 and 8 hours, respectively. NO(3) (-) also reduced the loss of nitrate reductase activity from mature root sections. In the presence of cycloheximide, both NADH-nitrate reductase and NO(3) (-)-induced NADH-cytochrome c reductase activities were lost at similar rates in root tips. NO(3) (-) protected the loss of NO(3) (-)-induced NADH-cytochrome c reductase to the same extent as that of NADH-nitrate reductase.

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PubMed1976

Effect of light and glucose on the induction of nitrate reductase and on the distribution of nitrate in etiolated barley leaves.

Barley seedlings grown in the dark with 10 mm KNO(3) have low levels of nitrate reductase activity even though large amounts of No(3) (-) accumulate in the leaves. When the leaves are excised and transferred to the light, there is an increase in nitrate reductase activity both in the presence and absence of exogenous NO(3) (-). When the leaves are transferred to a glucose solution (0.05 m) but kept in the dark, induction of nitrate reductase activity occurs only when fresh NO(3) (-) is added to the system.In dark-grown leaves, there are small traces of NO(3) (-) in a "metabolic pool." Addition of glucose does not alter this distribution. Light, on the other hand, results in an appreciable accumulation of NO(3) (-) in the metabolic pool. There is a linear correlation between nitrate reductase activity and the size of the metabolic NO(3) (-) pool. Our results thus suggest that NO(3) (-) accumulates in a storage pool when seedlings are grown in continuous darkness. The transfer of this NO(3) (-) to an active metabolic pool is mediated by light but not by glucose. We believe that this transfer of NO(3) (-) leads to the induction of nitrate reductase. When NO(3) (-) is included in the medium, both light and glucose increase its incorporation into the metabolic pool. The results suggest two mechanisms for regulating the metabolic NO(3) (-) pool: (a) a transfer from the storage pool which requires light; and (b) a transfer from the external medium which requires either glucose or light.

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PubMed1975

Effect of glucose on the induction of nitrate reductase in corn roots.

In Zea mays L., addition of glucose to the induction medium has no effect on the induction of nitrate reductase during the initial 3 hours either in root tips (0-10 mm) or mature root sections (25-35 mm). With longer times, higher levels of enzyme activity are recovered from both root segments when glucose is present in the incubation medium. The induction in root tips is saturated by 10 mm NO(3) (-). Higher concentrations of NO(3) (-) are required for saturation in mature root sections. The response to glucose is seen over a wide range of external NO(3) (-) concentrations.Nitrate reductase activity is lost rapidly when nitrate is withdrawn from the induction medium. Additions of glucose do not prevent this loss. Additions of glucose have no effect on total uptake of NO(3) (-) by the root segments but they increase the anaerobic NO(2) (-) production in both root tips and mature root segments. This latter measurement is considered to be an estimate of an active NO(3) (-) pool in the cytoplasm. Thus the results show that glucose alters the distribution of NO(3) (-) within the root sections. This may be an important factor in controlling the in vivo stability of the enzyme or its rate of synthesis.

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PubMed1974

Characteristics of an Acid protease from maize endosperm.

An assay has been developed to measure protease activity in endosperm extracts of maize seeds. With hemoglobin as substrate, the enzyme(s) has a pH optimum of 3.8 and a temperature optimum of 46 C. It also degrades gliadin, edestin, bovine serum albumin, and partially hydrolyzed zein and glutelin under standard assay conditions. The enzyme(s) has endopeptidase activity with all substrates tested. When undenatured zein and glutelin are suspended in an agar gel, both are efficiently degraded. Using this assay, the protease activity increases from day 3 to day 8 after inhibition and then declines.

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PubMed1974

The Hydrolysis of Endosperm Protein in Zea mays.

Degradation of the major storage proteins in maize endosperm, zein and glutelin, begins during the 2nd day of germination. The protein most abundant in the mature endosperm is degraded most rapidly. The patterns of protein loss are essentially similar in germinating seeds and excised endosperms. Cycloheximide, added at the beginning of the incubation period, prevents the development of alpha-amylase and protease activities and the disappearance of starch and protein reserves. Late additions (70 hours) of cycloheximide still inhibit the increase in hydrolase activity but have no effect on the hydrolysis of storage reserves. The results indicate that the hydrolytic enzymes are synthesized de novo in the maize endosperm.

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PubMed1972

Synthesis and turnover of nitrate reductase in corn roots.

THE INDUCTION AND REINDUCTION OF NITRATE REDUCTASE IN ROOT TIP OR MATURE ROOT SECTIONS SHOW ESSENTIALLY A SIMILAR PATTERN: a lag, a period of rapid increase in enzyme activity and finally a period of relatively minor change. Both inductions are sensitive to 6-methylpurine and cycloheximide. Kinetic studies with 6-methylpurine suggest that the half-life of the messenger RNA for nitrate reductase in both sections is about 20 minutes. The rate of decay of nitrate reductase activity induced by transfer to a nitrate-free medium is slower in root tips (t(1/2) = 3 hours) than in mature root sections (t(1/2) = 2 hours). The enzyme from mature root sections is also less stable to mild heat treatments (27 C; 40 C) than the enzyme from root tip sections. The results indicate that factors regulating enzyme turnover show important changes as root cells mature and may be significant in determining steady state levels of the enzyme.

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