Phosphorylation of phosphoenolpyruvate carboxykinase in plants. Studies in plants with C4 photosynthesis and Crassulacean acid metabolism and in germinating seeds

We have previously shown that phosphoenolpyruvate carboxykinase (PEPCK) is phosphorylated in vivo in the cotyledons of darkened cucumber seedlings and that phosphorylation is reversed by light [Walker and Leegood (1995) FEBS Lett. 362, 70–74]. In this study the molecular mass of PEPCK was estimated in a range of gluconeogenic seedlings and in leaves of C4 plants and plants with Crassulacean acid metabolism (CAM). Phosphorylation of PEPCK was studied in these plants by feeding tissues with [32P]Pi and assessing phosphorylation by SDS/PAGE and autoradiography of either total proteins or of immunoprecipitated protein. In gluconeogenic seedlings and most CAM plants PEPCK had a molecular mass of 74 kDa, whereas in C4 grasses the molecular mass of PEPCK was always smaller and varied from 67–71 kDa. In all gluconeogenic seedlings and leaves of CAM plants PEPCK was phosphorylated, but it was not phosphorylated in all species of C4 grasses studied. In CAM plants, phosphorylation of PEPCK occurred at night and dephosphorylation occurred during the day. In C4 grasses phosphorylation occurred when leaves were darkened and the enzyme was dephosphorylated following illumination, but it was only phosphorylated in those plants with larger (71 kDa) molecular mass forms of PEPCK

Effects of Phosphorylation on Phosphoenolpyruvate Carboxykinase from the C4 Plant Guinea Grass

In the C4 plant Guinea grass (Panicum maximum), phosphoenolpyruvate carboxykinase (PEPCK) is phosphorylated in darkened leaves and dephosphorylated in illuminated leaves. To determine whether the properties of phosphorylated and non-phosphorylated PEPCK were different, PEPCK was purified to homogeneity from both illuminated and darkened leaves. The final step of the purification procedure, gel filtration chromatography, further separated phosphorylated and non-phosphorylated forms. In the presence of a high ratio of ATP to ADP, the non-phosphorylated enzyme had a higher affinity for its substrates, oxaloacetate and phosphoenolpyruvate. The activity of the non-phosphorylated form was up to 6-fold higher when measured at low substrate concentrations. Comparison of proteoloytically cleaved PEPCK from Guinea grass, which lacked its N-terminal extension, from yeast (Saccharomyces cerevisiae), which does not possess an N-terminal extension, and from the C4 plant Urochloa panicoides, which possesses an N-terminal extension but is not subject to phosphorylation, revealed similar properties to the non-phosphorylated full-length form from Guinea grass. Assay of PEPCK activity in crude extracts of Guinea grass leaves, showed a large difference between illuminated and darkened leaves when measured in a selective assay (a low concentration of phosphoenolpyruvate and a high ratio of ATP to ADP), but there was no difference under assay conditions used to estimate maximum activity. Immunoblots of sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels showed no difference in the abundance of PEPCK protein in illuminated and darkened leaves. There were no light/dark differences in activity detected in maize (Zea mays) leaves, in which PEPCK is not subject to phosphorylation

Phosphoenolpyruvate Carboxykinase Assayed at Physiological Concentrations of Metal Ions Has a High Affinity for CO2

The effect of Mn2+/Mg2+ concentration on the activity of intact, homogeneous phosphoenolpyruvate carboxykinase (PEPCK) from leaves of the C4 grass, Guinea grass (Panicum maximum), have been investigated. Assay conditions were optimized so that PEPCK activity could be measured at concentrations of Mn2+/Mg2+ similar to those found in the cytosol (low micromolar Mn2+ and millimolar Mg2+). PEPCK activity was totally dependent on Mn2+ and was activated at low micromolar concentrations of Mn2+ by millimolar concentrations of Mg2+. Therefore, at physiological concentrations of Mn2+, PEPCK has a requirement for Mg2+. Assay at physiological concentrations of Mn2+/Mg2+ led to a marked decrease in its affinity for ATP and a 13-fold increase in its affinity for CO2. The Km (CO2) was further decreased by assay at physiological ATP to ADP ratios, reaching values as low as 20 μM CO2, comparable with the Km (CO2) of ribulose 1,5-bisphosphate carboxylase-oxygenase. This means that PEPCK will catalyze a reversible reaction and that it could operate as a carboxylase in vivo, a feature that could be particularly important in algal CO2-concentrating systems

Phosphoenolpyruvate Carboxykinase Is Involved in the Decarboxylation of Aspartate in the Bundle Sheath of Maize

We recently showed that maize (Zea mays L.) leaves contain appreciable amounts of phosphoenolpyruvate carboxykinase (PEPCK; R.P. Walker, R.M. Acheson, L.I. Técsi, R.C. Leegood [1997] Aust J Plant Physiol 24: 459–468). In the present study, we investigated the role of PEPCK in C4 photosynthesis in maize. PEPCK activity and protein were enriched in extracts from bundle-sheath (BS) strands compared with whole-leaf extracts. Decarboxylation of [4-14C]aspartate (Asp) by BS strands was dependent on the presence of 2-oxoglutarate and Mn2+, was stimulated by ATP, was inhibited by the PEPCK-specific inhibitor 3-mercaptopicolinic acid, and was independent of illumination. The principal product of Asp metabolism was phosphoenolpyruvate, whereas pyruvate was a minor product. Decarboxylation of [4-14C]malate was stimulated severalfold by Asp and 3-phosphoglycerate, was only slightly reduced in the absence of Mn2+ or in the presence of 3-mercaptopicolinic acid, and was light dependent. Our data show that decarboxylation of Asp and malate in BS cells of maize occurs via two different pathways: Whereas malate is mainly decarboxylated by NADP-malic enzyme, decarboxylation of Asp is dependent on the activity of PEPCK

Some thoughts of the future health of the South African Bantu

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Memorandum: what can be done to avoid coronary heart disease?

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Biological and disease patterns in South African inter-racial populations as modified by rise in privilege

Among the 4 ethnic groups in South Africa. populations may be observed in many stages of transition, from primitiveness to sopnistication. With the rise in socioeconomic circumstances, numerous changes have occurred. Those discussed include mortality rate and age structure, diet, growth, blood pressure, various biochemical measurements, physical activity, and disease pattern. The trend of changes implies that in the future, among non-Whites, diseases of nutritional inadequacy, especially protein-calorie malnutrition and pellagra, will decrease and will no longer be public health burdens. Simultaneously, however, there will be increases in conditions or diseases linked with nutritional excess, such as overweight and hypertension. In non-Whites, these and their ramifications are likely in t;me to exact the high tolls of mortality and morbidity from degenerative diseases which prevail in White populations. It is unlikely that recommendations directed at Whites and the affluent moieties of non-Whites will arrest the rising intensity of risk factors to health

Typhoid and Paratyphoid Fever in Children

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/67979/2/10.1177_000992286900800412.pd

Evidence for hadronic deconfinement in pˉ\bar{p}-p collisions at 1.8 TeV

We have measured deconfined hadronic volumes, $4.4 < V < 13.0$ fm$^{3}$, produced by a one dimensional (1D) expansion. These volumes are directly proportional to the charged particle pseudorapidity densities $6.75 < dN_{c}/d\eta < 20.2$. The hadronization temperature is $T = 179.5 \pm 5$ (syst) MeV. Using Bjorken's 1D model,the hadronization energy density is $\epsilon_{F} = 1.10 \pm 0.26$ (stat) GeV/fm$^{3}$ corresponding to an excitation of $24.8 \pm 6.2$ (stat) quark-gluon degrees of freedom.Comment: 15 pages, 3 figures, 2 table

Calculation of farfield distortion for a tilted-facet SOA

Semiconductor optical amplifiers (SOAs) are very important elements for telecommunications, computer communications, and signal processing applications. For stable, low noise operation, the modal reflection into the guided SOA mode must be minimized; modal reflectivity typically has to be kept below about {minus}40 dB. This can be accomplished by antireflection (AR) coatings, or by tilting of the SOA end facet. The latter approach has been vigorously pursued recently, because effective AR coatings require very high tolerances and have polarization-dependent reflectivities. Consequently, there has been a great deal of theoretical effort aimed at calculating the modal reflectivity from tilted interfaces, using a variety of approaches. However, there has been little attention directed toward calculating the transmitted field of a tilted-facet SOA. This is a problem of considerable importance, because the coupling of the SOA light to an element such as an optical fiber depends critically on the field distribution at the entrance plane to the fiber. Moreover, experimental measurements of the farfield of tilted-facet SOAs have revealed a curious crescent-shaped intensity distribution. To improve coupling efficiency it is important to understand to what extent this phenomenon is due to the SOA modal field distribution and to what extent it is due to the tilted interface. The authors explain the crescent-shaped farfield intensity distribution of tilted-facet SOAs using vector wave optics, and discuss implications for coupling to other optical elements