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Biomedical subjects

M Gibbs

Publications and source records attributed to M Gibbs.

At least 127 records · Page 7Linked to original sources
PubMed1975

H(2) metabolism in photosynthetic organisms: I. Dark h(2) evolution and uptake by algae and mosses.

Dark H(2) metabolism was studied in marine and fresh water red algae, the green alga, Chlamydomonas, and mosses. A time variable and temperature-sensitive anaerobic incubation was required prior to H(2) evolution. H(2) evolution was sensitive to disalicylidenepropanediamine. An immediate H(2) uptake was observed in these algae. Immediate dark H(2) uptake but no evolution was observed in the mosses.A cell-free hydrogenase preparation was obtained from anaerobically adapted Chlamydomonas reinhardii by means of sonic oscillation. The hydrogenase was not sedimented at 100,000g. It catalyzed the reduction of methylene blue, p-benzoquinone, NAD, NADP, but not spinach ferredoxin. H(2) evolution was noted with dithionite and with reduced methyl viologen as donors but not with reduced spinach ferredoxin. Similarly, hydrogenase activities were not affected by disalicylidenepropanediamine. The pH optima for H(2) evolution and for H(2) uptake were 7.2 and 7.5 to 9.5, respectively. Extracts prepared from the anaerobically adapted red alga, Chondrus crispus, and the moss, Leptobryum pyriforme, consumed but did not evolve H(2). Uptake was slightly stimulated by methylene blue. It is proposed that red algae and mosses appear to metabolize H(2) by a different pathway than Chlamydomonas.

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PubMed1974

Properties of phosphoribulokinase of whole chloroplasts.

The ability of intact spinach (Spinacia oleracea) chloroplast preparations to catalyze CO(2) fixation and photophosphorylation was examined. Under conditions optimal for CO(2) fixation, only poor photophosphorylation was observed. Conditions optimal for photophosphorylation were found to be highly inhibitory to the CO(2)-fixing capacity of the intact chloroplast preparation.A method for following the activity of phosphoribulokinase in the intact chloroplast preparation was developed, and conditions for optimal activity were defined. The enzyme was found to be activated 2- to 4-fold by preillumination with a half-time of less than 15 seconds. Activation was inhibited by magnesium ions and selectively by inhibitors of photosynthetic electron transport. We concluded that activation was due to the effect of a photoproduced reductant in a site preceding ferredoxin in the electron transport chain. The photoactivated state of the enzyme decayed in the dark with a half-time of about 8 minutes.

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PubMed1974

Carbon dioxide fixation in the light and in the dark by isolated spinach chloroplasts.

Factors affecting CO(2) fixation in the spinach (Spinacia oleracea) chloroplast were investigated. Free magnesium ions are shown to be highly inhibitory for photosynthetic CO(2) fixation in isolated intact spinach chloroplasts. The pH optimum for CO(2) fixation is about 8.5 but is dependent upon the reaction medium. Conditions are defined under which chloroplasts illuminated in the absence of CO(2) accumulate ribulose 1,5-diphosphate, and fix CO(2) in a subsequent dark period when high magnesium ion concentrations are provided. The regulation of photosynthetic CO(2) assimilation by these factors is discussed.

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PubMed1974

Photosynthetic intermediates, the warburg effect, and glycolate synthesis in isolated spinach chloroplasts.

Increasing levels of CO(2) have been shown to stimulate the rate of photosynthesis, eliminate the oxygen inhibition of photosynthesis (Warburg effect), and decrease glycolate formation in isolated spinach chloroplasts. Ribose 5-phosphate and fructose 1,6-diphosphate at concentrations of 5 to 10 mum also stimulate the rate of plastid photosynthesis and eliminate the Warburg effect. In contrast to the effect of high CO(2) levels, these sugar phosphates have little effect on glycolate formation. Evidence is presented to show that the level of intermediates of the photosynthetic carbon reduction cycle may influence the Warburg effect in vivo. It is postulated that the formation of glycolate is not the causal factor of the Warburg effect.

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PubMed1974

Comparative Enzymology of the Glyceraldehyde 3-Phosphate Dehydrogenases from Pisum sativum.

Glyceraldehyde 3-phosphate dehydrogenases (EC 1.2.1.12 and 1.2.1.13) have been purified from the seed, root, etiolated, and green shoot of peas (Pisum sativum). These enzymes are tetramers of 140,000 daltons, with subunits of 35,000 daltons. The enzymes differ in isoelectric point. The seed enzyme has a pI of 5.1, and the root enzyme has a pI of 4.5. The cytoplasmic enzyme from etiolated shoots is slightly acidic with a pI of 5.7 to 6.1 and is found in two separable forms. The chloroplast enzyme (from green shoots) is most basic with a pI of 8.0.In immunodiffusion experiments, the seed, root, and cytoplasmic enzymes of the etiolated shoot share antigenic homology, while the chloroplast enzyme does not cross react antigenically with the extra-chloroplast enzymes. The antiserum to the pea chloroplast enzyme did, however, cross react with glyceraldehyde 3-phosphate dehydrogenase purified from the spinach chloroplast. Therefore, the chloroplast enzyme is significantly different from the extra-chloroplast enzymes with respect to primary sequence.The NADP analog phosphoadenosine diphosphoribose showed competitive inhibition to the chloroplast enzyme with either pyridine nucleotide. The NAD analog pyridine 3-aldehyde NAD was competitive with respect to the NAD activity but was hyperbolic competitive in the presence of NADP, indicating a complexity in the binding of pyridine nucleotide to the chloroplast enzyme.

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PubMed1973

Nonreversible d-Glyceraldehyde 3-Phosphate Dehydrogenase of Plant Tissues.

Preparations of TPN-linked nonreversible d-glyceraldehyde 3-phosphate dehydrogenase (EC 1.2.1.9), free of TPN-linked reversible d-glyceraldehyde 3-phosphate dehydrogenase, have been obtained from green shoots, etiolated shoots, and cotyledons of pea (Pisum sativum), cotyledons of peanut (Arachis hypogea), and leaves of maize (Zea mays). The properties of the enzyme were similar from each of these sources: the Km values for d-glyceraldehyde 3-phosphate and TPN were about 20 mum and 3 mum, respectively. The enzyme activity was inhibited by l-glyceraldehyde 3-phosphate, d-erythrose 4-phosphate, and phosphohydroxypyruvate. Activity was found predominantly in photosynthetic and gluconeogenic tissues of higher plants. A light-induced, phytochrome-mediated increase of enzyme activity in a photosynthetic tissue (pea shoots) was demonstrated. Appearance of enzyme activity in a gluconeogenic tissue (endosperm of castor bean, Ricinus communis) coincided with the conversion of fat to carbohydrate during germination. In photosynthetic tissue, the enzyme is located outside the chloroplast, and at in vivo levels of triose-phosphates and pyridine nucleotides, the activity is probably greater than that of DPN-linked reversible d-glyceraldehyde 3-phosphate dehydrogenase. Several possible roles for the enzyme in plant carbohydrate metabolism are considered.

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PubMed1973

A mechanism for the indirect transfer of photosynthetically reduced nicotinamide adenine dinucleotide phosphate from chloroplasts to the cytoplasm.

A triose phosphate/3-phosphoglycerate shuttle for the indirect transfer of photosynthetically reduced NADP from chloroplasts to the cytoplasm has been demonstrated in vitro. Triose phosphate, formed from 3-phosphoglycerate in the chloroplast, was oxidized back to 3-phosphoglycerate outside the chloroplast by the nonreversible d-glyceraldehyde 3-phosphate dehydrogenase reaction which is specific for NADP. The 3-phosphoglycerate could presumably return to the chloroplast to complete the shuttle. The properties of nonreversible d-glyceraldehyde 3-phosphate dehydrogenase are considered particularly suitable for effective operation of this shuttle system.

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PubMed1972

Photosynthetic carbon metabolism of isolated corn chloroplasts.

Chloroplasts have been isolated from 4- to 6-day-old corn (Zea mays) leaves capable of assimilating 45 micromoles CO(2) per milligram chlorophyll per hour. The effects of various factors such as inorganic phosphate, reducing agents, inhibitors, intermediates of the photosynthetic carbon reduction cycle, organic acids, and oxygen on the photosynthetic rate and on the distribution of (14)C within the products by these chloroplasts were determined. The photosynthetic carbon metabolism of the corn plastids appeared to be similar to that already observed in spinach and pea chloroplasts. It was concluded that the corn plastids can fix CO(2) at meaningful rates via the photosynthetic carbon reduction cycle of Calvin without the operation of a cycle involving the C-4 compounds, malate and aspartate.

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PubMed1971

Formation of glycolate by a reconstituted spinach chloroplast preparation.

A reconstituted preparation requiring fructose 6-phosphate, transketolase, triphosphopyridine nucleotide, ferredoxin, fragmented spinach chloroplasts, and light capable of forming glycolate at rates of about 10 micromoles per milligram of chlorophyll per hour has been characterized. The glycolaldehyde-transketolase addition product could be substituted for fructose 6-phosphate and transketolase. The stoichiometry of the reaction was: 1 mole of fructose 6-phosphate consumed for each mole of glycolate and of reduced triphosphopyridine nucleotide produced. Evidence was presented indicating that glycolate formation was coupled to the photosystems of the photosynthetic electron transport chain. Synthesis of glycolate is envisaged as the result of either (a) a reaction between the upper two carbon atoms derived from fructose 6-phosphate and an uncharacterized oxidant generated by photosystem 2 or (b) hydrogen peroxide produced by the reoxidation of reduced triphos-phopyridine nucleotide or reduced ferredoxin by molecular oxygen.

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PubMed1971

Effect of antimycin a on photosynthesis of intact spinach chloroplasts.

Low concentrations (0.5-10 mum) of antimycin A were shown to increase the rate of CO(2) fixation, O(2) evolution and inorganic phosphate esterification in intact spinach (Spinacia oleracea) chloroplasts. The increase was highest when the light intensity was saturating. Stimulation was independent of the bicarbonate concentration and was accompanied by an enhancement in the synthesis of glycerate 3-phosphate with a decrease in dihydroxyacetone phosphate. The antibiotic decreased the Michaelis constant of the chloroplast but not of ribulose 1,5-diphosphate carboxylase for bicarbonate. It was suggested that antimycin A is affecting that portion (outer envelope) of the intact chloroplast which contains the enzyme mechanism for controlling the pace of CO(2) fixation.

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PubMed1971

Involvement of Photosynthetic Carbon Reduction Cycle Intermediates in CO(2) Fixation and O(2) Evolution by Isolated Chloroplasts.

The photosynthetic carbon reduction cycle intermediates can be divided into three classes according to their effects on the rate of photosynthetic CO(2) evolution by whole spinach (Spinacia oleracea) chloroplasts and on their ability to affect reversal of certain inhibitors (nigericin, arsenate, arsenite, iodoacetate, antimycin A) of photosynthesis: class I (maximal): fructose 1, 6-diphosphate, dihydroxyacetone phosphate, glyceraldehyde-3-phosphate, ribose-5-phosphate; class 2 (slight): glucose 6-phosphate, fructose 6-phosphate, ribulose-1, 5-diphosphate; class 3 (variable): glycerate 3-phosphate. While class 1 compounds influence the photosynthetic rate, they do not lower the Michaelis constant of the chloroplast for bicarbonate or affect strongly other photosynthetic properties such as the isotopic distribution pattern. It was concluded that the class 1 compounds influence the chloroplast by not only supplying components to the carbon cycle but also by activating or stabilizing a structural component of the chloroplast.

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PubMed1970

Glycolate formation in intact spinach chloroplasts.

Photosynthetic (14)CO(2) fixation and the accumulation of photosynthetic products and the response of each process to both 3-(3,4-dichlorophenyl)-1, 1-dimethylurea (DCMU) and ascorbate were investigated in the intact spinach chloroplast.Ascorbate increased the rate of CO(2) uptake with an increase in all photosynthetic products, but, proportionally, there was a much larger increase in glycolate formation. CO(2) fixation inhibited by DCMU was partially restored on addition of ascorbate. Under conditions not optimal for glycolate formation, such as saturating levels of CO(2) and an anaerobic atmosphere, ascorbate in the presence of DCMU restored the formation of all photosynthetic products excluding glycolate. This effect of ascorbate on glycolate synthesis in the presence of DCMU was diminished under conditions which favored glycolate formation. Externally added glycerate 3-phosphate and fructose 1,6-diphosphate depressed the appearance of radioactivity in glycolate.The data are interpreted to indicate that glycolate is produced during photosynthesis as a result of a reaction between a 2-carbon piece derived from a sugar phosphate and an oxidant generated by the photochemical act. The oxidant may be an intermediate of photosystem 2 or a peroxide generated by a mechanism of the Mehler type involving molecular oxygen.

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