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1.
Evert , R. F. (U. Wisconsin, Madison.) The cambium and seasonal development of the phloem in Pyrus malus. Amer. Jour. Bot. 50(2): 149–159. Illus. 1963.—The cambium in apple consists of several layers of cells at all times, and practically all cambial cells divide periclinally one or more times before undergoing differentiation. The cambial initials do not seem to be in a uniform, uniseriate layer. Judged by collections made during 2 seasons (August, 1958–October, 1960), the seasonal cycle of phloem development is as follows. Early in April, cells in the outer margin of the cambial zone begin to differentiate into sieve elements. At approximately the same time, activity (division) commences throughout the cambial zone. By the end of July or early August, sieve-element differentiation is completed. Cessation of function begins in either late September or in October with the formation of definitive callose on the sieve areas of sieve elements in the outer margin of the functional phloem. By late November, all sieve elements are devoid of contents and most of their companion cells collapsed. Phloem differentiation precedes xylem differentiation by approximately a month and a half; xylem and phloem differentiation cease almost simultaneously; and fiber-sclereid development is coincident with the period of maximal xylem differentiation.  相似文献   

2.
Slices of hypocotyls from 3-day-old seedlings of Vigna sesquipedalis (L.) Fruwirth in the germination stage were incubated under various gaseous conditions. The NADP+NADPH level in the hypocotyl slices changed with the oxygen tension. A high NADP+NADPH level was observed under aerobic conditions and a low NADP+NADPH level under anaerobic conditions.

The 100 × NADH/NAD+NADH ratio increased greatly under anaerobic conditions. In general a low NADP + NADPH level corresponded with a high 100 × NADH/NAD+NADH ratio. On the basis of the results given in the following paper, it was discussed that the slowness of NADH oxidation in hypocotyl tissue due to anaerobic conditions results in the inhibition of NADP formation.

The variation of the NADP+NADPH level was considered to produce a modification of the carbohydrate metabolism.

The NADP+NADPH level in E. coli cells suspended in glucose solution also changed with the oxygen tension.

  相似文献   

3.
Properties of glutamate dehydrogenase purified from Bacteroides fragilis   总被引:2,自引:0,他引:2  
The dual pyridine nucleotide-specific glutamate dehydrogenase [EC 1.4.1.3] was purified 37-fold from Bacteroides fragilis by ammonium sulfate fractionation, DEAE-Sephadex A-25 chromatography twice, and gel filtration on Sephacryl S-300. The enzyme had a molecular weight of approximately 300,000, and polymeric forms (molecular weights of 590,000 and 920,000) were observed in small amounts on polyacrylamide gel disc electrophoresis. The molecular weight of the subunit was 48,000. The isoelectric point of the enzyme was pH 5.1. This glutamate dehydrogenase utilized NAD(P)H and NAD(P)+ as coenzymes and showed maximal activities at pH 8.0 and 7.4 for the amination with NADPH and with NADH, respectively, and at pH 9.5 and 9.0 for the deamination with NADP+ and NAD+, respectively. The amination activity with NADPH was about 5-fold higher than that with NADH. The Lineweaver-Burk plot for ammonia showed two straight lines in the NADPH-dependent reactions. The values of Km for substrates were: 1.7 and 5.1 mM for ammonium chloride, 0.14 mM for 2-oxoglutarate, 0.013 mM for NADPH, 2.4 mM for L-glutamate, and 0.019 mM for NADP+ in NADP-linked reactions, and 4.9 mM for ammonium chloride, 7.1 mM for 2-oxoglutarate, 0.2 mM for NADH, 7.3 mM for L-glutamate, and 3.0 mM for NAD+ in NAD-linked reactions. 2-Oxoglutarate and L-glutamate caused substrate inhibition in the NADPH- and NADP+-dependent reactions, respectively, to some extent. NAD+- and NADH-dependent activities were inhibited by 50% by 0.1 M NaCl. Adenine nucleotides and dicarboxylic acids did not show remarkable effects on the enzyme activities.  相似文献   

4.
The yeast Candida parapsilosis possesses two routes of electron transfer from exogenous NAD(P)H to oxygen. Electrons are transferred either to the classical cytochrome pathway at the level of ubiquinone through an NAD(P)H dehydrogenase, or to an alternative pathway at the level of cytochrome c through another NAD(P)H dehydrogenase which is insensitive to antimycin A. Analyses of mitoplasts obtained by digitonin/osmotic shock treatment of mitochondria purified on a sucrose gradient indicated that the NADH and NADPH dehydrogenases serving the alternative route were located on the mitochondrial inner membrane. The dehydrogenases could be differentiated by their pH optima and their sensitivity to amytal, butanedione and mersalyl. No transhydrogenase activity occurred between the dehydrogenases, although NADH oxidation was inhibited by NADP+ and butanedione. Studies of the effect of NADP+ on NADH oxidation showed that the NADH:ubiquinone oxidoreductase had Michaelis-Menten kinetics and was inhibited by NADP+, whereas the alternative NADH dehydrogenase had allosteric properties (NADH is a negative effector and is displaced from its regulatory site by NAD+ or NADP+).  相似文献   

5.
The interactions of calcium with NAD+, NADH, NADP+ and NADPH in a 50% (by volume) methanol/water mixture (pH 7, 25 degrees C) were studied by calorimetry. The association constants for 1:1 complex formation were found to be 6.6 +/- 0.2, 270 +/- 76, 18 +/- 3 and 98 +/- 10 for NAD+, NADH, NADP+ and NADPH, respectively. Comparing these to the association constants for an aqueous system reveals that as the polarity of the solvent system is decreased the interactions involving NAD+, NADP+ and NADPH are all decreased. In contrast, the interaction involving NADH is markedly increased. All the interactions were found to be endothermic.  相似文献   

6.
The seasonal development of phloem in the stems of Siberian larch (Larix sibirica Ldb.) was studied over two seasons on 50–60-year-old trees growing in a natural stand in the Siberian forest-steppe zone. Trees at the age of 20–25 years were used to study metabolites in differentiating and mature phloem elements, cambial zone, and radially growing xylem cells in the periods of early and late wood formation. The development of the current-year phloem in the stems of 50–60-year-old trees started, depending on climatic conditions, in the second-third decades of May, 10–20 days before the xylem formation, and ended together with the shoot growth cessation in late July. Monitoring of the seasonal activity of cambium producing phloem sieve cells and the duration of their differentiation compared to the xylem derivatives in the cambium demonstrated that the top production of phloem and xylem cells could coincide or not coincide during the season, while their differentiation activity was always in antiphase. Sieve cells in the early phloem are separated from those in the late phloem by a layer of tannin-containing cells, which are formed in the period when late xylem formation starts. The starch content in the structural elements of phloem depends on the state of annual xylem layer development. The content of low molecular weight carbohydrates, amino acids, organic acids, and phenols in phloem cells, cambial zone, and xylem derivatives of the cambium depends on the cell type and developmental stage as well as on the type of forming wood (early or late) differing by the cell wall parameters and, hence, by the requirement for assimilates. Significant differences in the dynamics of substances per dry weight and cell were observed during cell development.  相似文献   

7.
Coenzyme specificity of mammalian liver D-glycerate dehydrogenase   总被引:1,自引:0,他引:1  
D-Glycerate dehydrogenase (glyoxylate reductase) was partially purified from rat liver by anion- and cation-exchange chromatography. When assayed in the direction of D-glycerate or glycolate formation, the enzyme was inhibited by high (greater than or equal to 0.5 mM), unphysiological concentrations of hydroxypyruvate or glyoxylate much more potently in the presence of NADPH than in the presence of NADH. However, the dehydrogenase displayed a much greater affinity for NADPH (Km less than 1 microM) than for NADH (Km = 48-153 microM). Furthermore, NADP was over 1000-fold more potent than NAD in inhibiting the enzyme competitively with respect to NADH. NADP also inhibited the reaction competitively with respect to NADPH whereas NAD, at concentrations of up to 10 mM had no inhibitory effect. When measured by the formation of hydroxypyruvate from D-glycerate, the enzyme also displayed a much greater affinity for NADP than for NAD. These properties indicate that liver D-glycerate dehydrogenase functions physiologically as an NADPH-specific reductase. In agreement with this conclusion, the addition of hydroxypyruvate or glyoxylate to suspensions of rat hepatocytes stimulated the pentose-phosphate pathway. The coenzyme specificity of D-glycerate dehydrogenase is discussed in relation to the biochemical findings made in D-glyceric aciduria and in primary hyperoxaluria type II (L-glyceric aciduria).  相似文献   

8.
The increase of sorbitol and fructose levels caused by aldose reductase activation and sorbitol dehydrogenase inhibition were observed in sciatic nerve of streptozotocin-diabetic rats. Elevated polyol pathway activity has been implicated in the development of diabetic complications such as neuropathy. The regulation of polyol pathway enzymes is based on the changes of redox state of free nicotinamide nucleotides. The decrease of the NADP+/NADPH ratio in cytosolic compartment of sciatic nerve cells activated aldose reductase and the decrease of the NAD+/NADH ratio inhibited sorbitol dehydrogenase. Nicotinamide as a precursor of NAD+ biosynthesis increased the free NADP+/NADPH and NAD+/NADH ratios and inhibited the activity of polyol pathway. The sorbitol level decreased in sciatic nerve of nicotinamide-treated streptozotocin-diabetic rats as compared to non-treated ones. Thus, the data provide evidence for important role of nicotinamide, as an antidiabetic drug, in prevention or correction of diabetic neuropathy.  相似文献   

9.
The goal was to determine whether endogenous cytosolic NAD(P)H:quinone oxidoreductase 1 (NQO1) preferentially uses NADPH or NADH in intact pulmonary arterial endothelial cells in culture. The approach was to manipulate the redox status of the NADH/NAD(+) and NADPH/NADP(+) redox pairs in the cytosolic compartment using treatment conditions targeting glycolysis and the pentose phosphate pathway alone or with lactate, and to evaluate the impact on the intact cell NQO1 activity. Cells were treated with 2-deoxyglucose, iodoacetate, or epiandrosterone in the absence or presence of lactate, NQO1 activity was measured in intact cells using duroquinone as the electron acceptor, and pyridine nucleotide redox status was measured in total cell KOH extracts by high-performance liquid chromatography. 2-Deoxyglucose decreased NADH/NAD(+) and NADPH/NADP(+) ratios by 59 and 50%, respectively, and intact cell NQO1 activity by 74%; lactate restored NADH/NAD(+), but not NADPH/NADP(+) or NQO1 activity. Iodoacetate decreased NADH/NAD(+) but had no detectable effect on NADPH/NADP(+) or NQO1 activity. Epiandrosterone decreased NQO1 activity by 67%, and although epiandrosterone alone did not alter the NADPH/NADP(+) or NADH/NAD(+) ratio, when the NQO1 electron acceptor duroquinone was also present, NADPH/NADP(+) decreased by 84% with no impact on NADH/NAD(+). Duroquinone alone also decreased NADPH/NADP(+) but not NADH/NAD(+). The results suggest that NQO1 activity is more tightly coupled to the redox status of the NADPH/NADP(+) than NADH/NAD(+) redox pair, and that NADPH is the endogenous NQO1 electron donor. Parallel studies of pulmonary endothelial transplasma membrane electron transport (TPMET), another redox process that draws reducing equivalents from the cytosol, confirmed previous observations of a correlation with the NADH/NAD(+) ratio.  相似文献   

10.
Functional sieve cells are present at all times in the secondary phloem of Pinus banksiana Lamb., P. resinosa Ait., and P. strobus L. With regard to a given year's growth increment, all but the last-formed sieve cells (2-4 layers) cease functioning the same season they are derived from the cambium. The former overwinter and remain functional until new sieve cells differentiate in spring. Toward the end of March undifferentiated cells in the outer margin of the cambial zone begin to differentiate into sieve cells. About a week later, cambial activity (cell division) commences. All early phloem is produced by early May before new xylem differentiation begins. Most sieve cells are differentiated by late August, but a few not until late September. Cessation of function begins in late May or June with formation of definitive callose on sieve areas of the sieve cells which overwintered and continues slowly to sieve cells of the current season's early phloem. By mid-December all but the last-formed sieve cells (i.e., those which will overwinter in a functional state) are devoid of contents. Phloem differentiation precedes xylem differentiation by approximately 1 1/2 months. Xylem and phloem production cease more or less simultaneously in August, xylem and phloem differentiation in September.  相似文献   

11.
Circular patches of bark were surgically isolated on the sides of trembling aspen (Populus tremuloides Michx.) trees at breast height at various times during the dormant and growing seasons. Subsequently, samples of wood and attached bark were taken from isolated and control sites to determine the effects of isolation of the bark on cambial activity and xylem and phloem development. In control trees cambial activity and xylem and phloem development occurred normally. Isolation of bark during the dormant season (in November, February, or March) did not prevent initiation of cambial activity and of phloem differentiation in spring but continued normal cambial activity and phloem developmented were prevent. Xylem differentiation was essentially prevented by isolation of tissues during the dormant season. The ultimate effect of isolation of the bark on the cambium, either during the dormant season or during the growing season, was subdivision of all fusiform cambial cells into strands of parenchymatous elements; the ultimate effect on the newly formed phloem was early death of the sieve elements. The most conspicuous effect of isolation of the bark after xylem differentiation had begun was the curtailment of secondary wall formation. Shortening of cells of the cambial region was reflected in the length of the vessel members which differentiated from such cells. These results indicate that normal cambial activity and xylem and phloem development require a supply of currently translocated regulatory substances from the shoots.  相似文献   

12.
Circular patches of bark were surgically isolated on the sides of sugar maple (Acer saccharum Marsh.) trees at breast height at various times during the dormant and growing seasons. Subsequently, samples of wood and attached bark were taken from isolated and control sites to determine the effects of isolation of the bark on cambial activity and xylem and phloem development. In control sites cambial activity and xylem and phloem development occurred normally. Isolation of bark during the dormant season (in November, February, or March) prevented initiation of cambial activity and xylem and phloem development in isolated areas of half of the trees. Varying degrees of cambial activity (periclinal divisions) occurred in the remaining isolated areas, but normal cambial activity and xylem and phloem development were prevented. Isolation of bark after initiation of cambial activity and phloem differentiation, but prior to initiation of xylem differentiation, resulted in the formation of very narrow xylem and phloem increments with atypically short vessel members and sieve-tube members, respectively. The xylem increments consisted primarily of parenchyma cells. Isolation of bark after initiation of xylem differentiation resulted in curtailment of secondary wall formation in the last-formed part of many increments. The last-formed vessel members of all these xylem increments were atypically short. Similarly, the last formed sieve-tube members of corresponding phloem increments were atypically short. The atypically short cells in the xylem and phloem of isolated areas reflected the effect of isolation on the cambial region, viz., the subdivision of all fusiform cells into strands of cells. Ultimately, the strands of short fusiform cells lapsed into maturity, leaving only strands of parenchymatous elements between xylem and phloem.  相似文献   

13.
When the extracellular concentration of glucose was raised from 3 mM to 7 mM (the concentration interval in which beta-cell depolarization and the major decrease in K+ permeability occur), the cytosolic free [NADPH]/[NADP+] ratio in mouse pancreatic islets increased by 29.5%. When glucose was increased to 20 mM, a 117% increase was observed. Glucose had no effect on the cytosolic free [NADH]/[NAD+] ratio. Neither the cytosolic free [NADPH]/[NADP+] ratio nor the corresponding [NADH]/[NAD+] ratio was affected when the islets were incubated with 20 mM-fructose or with 3 mM-glucose + 20 mM-fructose, although the last-mentioned condition stimulated insulin release. The insulin secretagogue leucine (10 mM) stimulated insulin secretion, but lowered the cytosolic free [NADPH]/[NADP+] ratio; 10 mM-leucine + 10 mM-glutamine stimulated insulin release and significantly enhanced both the [NADPH]/[NADP+] ratio and the [NADH]/[NAD+] ratio. It is concluded that the cytosolic free [NADPH]/[NADP+] ratio may be involved in coupling beta-cell glucose metabolism to beta-cell depolarization and ensuing insulin secretion, but it may not be the sole or major coupling factor in nutrient-induced stimulation of insulin secretion.  相似文献   

14.
Abstract Extracts from the obligate methylotroph Methylobacillus flagellatum KT and its temperature-sensitive (ts) glucose 6-phosphate dehydrogenase (GPD) mutants were analysed by electrophoresis, isoelectrofocusing and chromatography methods. GPD is present in two forms differing in the isoelectric point (IEP) values, but identical in other properties. Both forms are specific to NAD and NADP, have similar affinity to substrates, exhibit equal levels of inhibition by NAD(P)H and ATP and have the same dependence of activity on temperature. The synthesis of both forms is controlled by one gene. 6-phosphogluconate dehydrogenase (GND) is represented by two proteins with different IEP values. One is specific both to NAD and NADP, is stable and inhibited by NADH and NADPH to a similar extent. The second is specific to NAD only, unstable and inhibited by NADH to a greater extent than by NADPH.  相似文献   

15.
Initial-rate studies were made of the oxidation of L-glutamate by NAD+ and NADP+ catalysed by highly purified preparations of dogfish liver glutamate dehydrogenase. With NAD+ as coenzyme the kinetics show the same features of coenzyme activation as seen with the bovine liver enzyme [Engel & Dalziel (1969) Biochem. J. 115, 621--631]. With NADP+ as coenzyme, initial rates are much slower than with NAD+, and Lineweaver--Burk plots are linear over extended ranges of substrate and coenzyme concentration. Stopped-flow studies with NADP+ as coenzyme give no evidence for the accumulation of significant concentrations of NADPH-containing complexes with the enzyme in the steady state. Protection studies against inactivation by pyridoxal 5'-phosphate indicate that NAD+ and NADP+ give the same degree of protection in the presence of sodium glutarate. The results are used to deduce information about the mechanism of glutamate oxidation by the enzyme. Initial-rate studies of the reductive amination of 2-oxoglutarate by NADH and NADPH catalysed by dogfish liver glutamate dehydrogenase showed that the kinetic features of the reaction are very similar with both coenzymes, but reactions with NADH are much faster. The data show that a number of possible mechanisms for the reaction may be discarded, including the compulsory mechanism (previously proposed for the enzyme) in which the sequence of binding is NAD(P)H, NH4+ and 2-oxoglutarate. The kinetic data suggest either a rapid-equilibrium random mechanism or the compulsory mechanism with the binding sequence NH4+, NAD(P)H, 2-oxoglutarate. However, binding studies and protection studies indicate that coenzyme and 2-oxoglutarate do bind to the free enzyme.  相似文献   

16.
In Escherichia coli, the pentose phosphate pathway is one of the main sources of NADPH. The first enzyme of the pathway, glucose-6-phosphate dehydrogenase (G6PDH), is generally considered an exclusive NADPH producer, but a rigorous assessment of cofactor preference has yet to be reported. In this work, the specificity constants for NADP and NAD for G6PDH were determined using a pure enzyme preparation. Absence of the phosphate group on the cofactor leads to a 410-fold reduction in the performance of the enzyme. Furthermore, the contribution of the phosphate group to binding of the transition state to the active site was calculated to be 3.6 kcal·mol(-1). In order to estimate the main kinetic parameters for NAD(P) and NAD(P)H, we used the classical initial-rates approach, together with an analysis of reaction time courses. To achieve this, we developed a new analytical solution to the integrated Michaelis-Menten equation by including the effect of competitive product inhibition using the ω-function. With reference to relevant kinetic parameters and intracellular metabolite concentrations reported by others, we modeled the sensitivity of reduced cofactor production by G6PDH as a function of the redox ratios of NAD/NADH (rR(NAD)) and NADP/NADPH (rR(NADP)). Our analysis shows that NADPH production sharply increases within the range of thermodynamically feasible values of rR(NADP), but NADH production remains low within the range feasible for rR(NAD). Nevertheless, we show that certain combinations of rR(NADP) and rR(NAD) sustain greater levels of NADH production over NADPH.  相似文献   

17.
The fate of all aerobic organisms is dependent on the varying intracellular concentrations of NADH and NADPH. The former is the primary ingredient that fuels ATP production via oxidative phosphorylation, while the latter helps maintain the reductive environment necessary for this process and other cellular activities. In this study we demonstrate a metabolic network promoting NADPH production and limiting NADH synthesis as a consequence of an oxidative insult. The activity and expression of glucose-6-phosphate dehydrogenase, malic enzyme, and NADP(+)-isocitrate dehydrogenase, the main generators of NADPH, were markedly increased during oxidative challenge. On the other hand, numerous tricarboxylic acid cycle enzymes that supply the bulk of intracellular NADH were significantly downregulated. These metabolic pathways were further modulated by NAD(+) kinase (NADK) and NADP(+) phosphatase (NADPase), enzymes known to regulate the levels of NAD(+) and NADP(+). While in menadione-challenged cells, the former enzyme was upregulated, the phosphatase activity was markedly increased in control cells. Thus, NADK and NADPase play a pivotal role in controlling the cross talk between metabolic networks that produce NADH and NADPH and are integral components of the mechanism involved in fending off oxidative stress.  相似文献   

18.
1. The concentrations of the nicotinamide-adenine dinucleotides in rat liver have been determined at intervals during the period 1-24hr. after feeding adult female rats with dimethylnitrosamine or thioacetamide. 2. The administration of dimethylnitrosamine resulted in a rapid decrease in the sum of NAD+NADH(2). This sum was decreased by 40% 3hr. after dosing. 3. Dimethylnitrosamine administration also produced an overall decrease in the NADP+NADPH(2) but this decrease was not so early nor as marked as that found for NAD+NADH(2). 4. The changes produced by thioacetamide were quite different from those obtained with dimethylnitrosamine. Thioacetamide produced a temporary rise in the NAD+NADH(2) followed by a small fall. The NADP+NADPH(2) was little changed in the early hours after dosing with thioacetamide but had decreased by approx. 15% 18hr. after administration. 5. These changes are discussed in terms of the known hepatotoxic actions of dimethylnitrosamine and thioacetamide, and are compared with previously reported changes found after the administration of carbon tetrachloride.  相似文献   

19.
The effects of coenzymes NAD(P) and NAD(P)H on the kinetics of the ox liver glutamate dehydrogenase reaction have been studied. The oxidized coenzymes were shown to activate alpha-ketoglutarate amination at inhibiting concentrations of NADH and NADPH. The reduced coenzymes, NADH and NADPH, inhibit glutamate deamination with both NAD and NADP as coenzymes. The data obtained are discussed in terms of literature data on the mechanisms of the coenzyme effects on the glutamate dehydrogenase activity and are inconsistent with the theory of direct ligand--ligand interactions. It was shown that the peculiarities of the glutamate dehydrogenase kinetics can easily be interpreted in the light of the two state models.  相似文献   

20.
The cambium in black locust consists of several layers of cells at all times. Cambial reactivation (division) is preceded by a decrease in density of cambial cell protoplasts and cell wall thickening but not by cell enlargement. During the resumption of cambial activity, periclinal divisions occur throughout the cambial zone. Early divisions contribute largely to the phloem side. The period of greatest cambial activity coincides with early wood formation. Judged by numerous collections made during two seasons (October, 1960-October, 1962) the seasonal cycle of phloem development is as follows. Phloem differentiation begins in early April, ends in late September. The amount of phloem produced is quite variable (range: 1-10 bands of sieve elements per year). Cessation of function begins with the accumulation of definitive callose in the first-formed sieve elements and spreads to those more recently formed. By late November all but the last-formed sieve elements are collapsed. All sieve elements are collapsed by mid-winter and before the resumption of new phloem production in spring. Phloem differentiation precedes xylem differentiation by at least 1 week, and apparently functional sieve elements are present 3 weeks before new functional vessel elements. Xylem and phloem production ends simultaneously in most trees.  相似文献   

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