Enzyme which catalyses the phosphorylation of adp

Clicking on the donut icon will load a page at altmetric. Find more information on the Altmetric Attention Score and how the score is calculated. Ambitions to biomanufacture sulfate-containing compounds such as heparin and chondroitin sulfate also promote the study on PAPS in vitro synthesis. The efficient enzymatic route that is constructed here for PAPS synthesis with low cost would boost the biosynthesis of sulfated compounds and peptides.

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More by Ruirui Xu. More by Yang Wang. Plant Mol. Adenosine diphosphate glucose pyrophosphorylase genes in wheat: differential expression and gene mapping. Shifting the limits in wheat research and breeding using a fully annotated reference genome. Heat stability of the potato tuber ADP-glucose pyrophosphorylase: role of Cys residue 12 in the small subunit.

ADP-glucose pyrophosphorylase, a regulatory enzyme for bacterial glycogen synthesis. ADP-glucose pyrophosphorylase: A regulatory enzyme for plant starch synthesis. Resurrecting the ancestral enzymatic role of a modulatory subunit.

Investigation of the interaction between the large and small subunits of potato ADP-glucose pyrophosphorylase. PloS Comput. A spatiotemporal analysis of enzymatic activities associated with carbon metabolism in wild-type and mutant embryos of Arabidopsis using in situ histochemistry. Plant J. Non-phosphorylating glyceraldehydephosphate dehydrogenase is post-translationally phosphorylated in heterotrophic cells of wheat Triticum aestivum. FEBS Lett. Formation of oil in the seed of Ricinus communis L.

The different large subunit isoforms of Arabidopsis thaliana ADP-glucose pyrophosphorylase confer distinct kinetic and regulatory properties to the heterotetrameric enzyme. Critical and speculative review of the roles of multi-protein complexes in starch biosynthesis in cereals. Plant Sci. Cross-phosphorylation between Arabidopsis thaliana sucrose nonfermenting 1-related protein kinase 1 AtSnRK1 and its activating kinase AtSnAK determines their catalytic activities.

Activities of enzymes involved in starch synthesis in wheat grains differing in starch content. Plant Physiol. On the roles of wheat endosperm ADP-glucose pyrophosphorylase subunits. A simple method for the isolation and purification of total lipids from animal tissues.

A colorimetric method for the assay of ADP-glucose pyrophosphorylase. ADP-glucose pyrophosphorylase from wheat endosperm. Purification and characterization of an enzyme with novel regulatory properties. Planta , — Regulation of starch biosynthesis in response to a fluctuating environment. Duplications and functional divergence of ADP-glucose pyrophosphorylase genes in plants. BMC Evol.

Phylogenetic analysis of ADP-glucose pyrophosphorylase subunits reveals a role of subunit interfaces in the allosteric properties of the enzyme. Food security: The challenge of feeding 9 billion people.

Science , — Starch formation inside plastids of higher plants. Protoplasma , — Seed development in Ricinus communis castor bean I. Descriptive morphology. Plant Cell 13 , — Starch biosynthesis in cereal endosperm. Regulation of starch metabolism: The age of enlightenment? Plant Biol. Investigation of subunit function in ADP-glucose pyrophosphorylase.

Advances in Phos-tag-based methodologies for separation and detection of the phosphoproteome. Acta - Proteins Proteomics , — Insensitivity of barley endosperm ADP-glucose pyrophosphorylase to 3-phosphoglycerate and orthophosphate regulation.

Ostreococcus tauri ADP-glucose pyrophosphorylase reveals alternative paths for the evolution of subunit roles. The ancestral activation promiscuity of ADP-glucose pyrophosphorylases from oxygenic photosynthetic organisms. Efficient site-directed mutagenesis using uracil-containing DNA.

Methods Enzymol. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature , — Heat stability of maize endosperm ADP-glucose pyrophosphorylase is enhanced by insertion of a cysteine in the N terminus of the small subunit.

Phosphorylation site mapping of soluble proteins: Bioinformatical filtering reveals potential plastidic phosphoproteins in Arabidopsis thaliana. BMC Genomics 15 , 1— Starch as a source, starch as a sink: The bifunctional role of starch in carbon allocation.

Towards the rational design of cereal starches. Selective adsorption of phosphoproteins on gel-immobilized ferric chelate. Biochemistry 25 , — Proteomic and morphological analysis of early stages of wheat grain development.

Proteomics 10 , — Proteomic profile of the nucellus of castor bean Ricinus communis L. Proteomics 75 , — Different isoforms of starch-synthesizing enzymes controlling amylose and amylopectin content in rice Oryza sativa L. Purification and properties of nonproteolytic degraded ADP-glucose pyrophosphorylase from maize endosperm. Several polyphenolic phytochemicals, such as quercetin and resveratrol, have been known to affect the activity ATPase. At decreased concentrations, it inhibits both soluble and insoluble mitochondrial ATPase.

However, it does not impact oxidative phosphorylation occurring in other mitochondrial entities [ 39 , 40 , 41 ]. This scheme is based on the binding change mechanism of ATP hydrolysis [ 36 ]. IF1 is a naturally occurring 9. Several other plant products also serve as ATPase inhibitors. Polyphenols and flavones has been found effective in the inhibition of bovine and porcine heart F 0 F 1 -ATPase [ 41 , 42 ].

Efrapeptins are peptides which are produced by fungi of the genus Tolypocladium that have antifungal, insecticidal and mitochondrial ATPase inhibitory activities [ 43 ]. The mode of inhibition is competitive with ADP and phosphate [ 30 ].

Another inhibitor piceatannol, a stilbenoid, has been found to inhibit the F-type ATPase preferably by targeting the F 1 subunit [ 39 ]. Another inhibitor of ATPase is bicarbonate. Bicarbonate anion acts as activator of ATP hydrolysis and Lodeyro et. This inhibition of ATP synthase activity was competitive with respect to ADP at low fixed phosphate concentration, mixed at high phosphate concentration and non-competitive towards Pi at any fixed ADP concentration [ 44 ].

Other inhibitors of ATPase are tenoxin, lecucinostatin, fluro-aluminate, dicyclohexyl-carbodimide and azide. Leucinostatins bind to the F 0 part of ATP synthases and inhibit oxidative phosphorylation in mitochondria and photophosphorylation in chloroplasts [ 46 ].

Dicyclohexylcarbodiimide DCCD reacts with the carboxyl group of the conserved acidic amino acid residue of subunit c at higher pH levels. So this compound can be considered as an inhibitor of both F O and F 1. However, inhibition of F O is highly specific, well-defined, and requires a much lower concentration of the inhibitor [ 48 ]. The list of inhibitors that directly and indirectly inhibit the activity of ATP synthase includes, magnesium, bismuth subcitrate and omeprazole, ethidium bromide, adenylyl imidodiphosphate, arsenate, angiostatin and enterostatin, ossamycin, dequalinium and methionine, almitrine, apoptolidin, aurovertin and citreoviridin, rhodamines, venturicidin, estrogens, catechins, kaempferol, genistein, biochanin A, daidzein and continues to grow [ 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , 61 , 62 ].

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Mitochondrial disorders. The Scientific World Journal. Published Jan ATP synthase a marvellous rotary engine of the cell. Nature Reviews Molecular Cell Biology. Mitochondrial ATP synthase: architecture, function and pathology. Journal of Inherited Metabolic Disease. Organization and Regulation of Mitochondrial Oxidative Phosphorylation.

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Web site. Accessed March 26, Journal of Experimental Biology. The rotary machine in the cell, ATP synthase. Journal of Biological Chemistry. R Search in Google Scholar. Accompanying seed maturation and starch accumulation, a combined regulation of ADP-Glc PPase by metabolites and phosphorylation may provide an enzyme with stable levels of activity.

Such concerted modulation would drive carbon skeletons to the synthesis of starch for its long-term storage, which later support seed germination. Starch is a major product of photosynthesis performed by vascular plants and constitutes the foremost storage of carbon and energy in grasses Jeon et al.

Cereals wheat, maize, barley, and rice as more relevant in production store starch in seed endosperm, and they supply more than half of the caloric demands in the world population Tuncel and Okita, ; Goren et al. Enhanced biosynthesis of the polysaccharide greatly influences the grain yield of cereals having starch as the principal reserve compound Jeon et al. Improvement in quantity and quality in the production of key harvestable grains is a challenge to be solved in the coming decades.

Indeed, it is projected a critical demographic expansion together with increasing industrial requirements of feedstock for biofuels, bioplastics, and bioadhesives to cope with climate change by the mid-century Morell and Myers, ; Godfray et al. In this scenario, the in-depth understanding of the process of starch biosynthesis is relevant to better design strategies to increase yields in plants of agronomic interest.

Starch accumulates in plastids of both photosynthetic and non-photosynthetic plant cells Jeon et al. The metabolic route follows by the action of enzymes implicated in branching, debranching, phosphorylation, and de-phosphorylation of the polymer under formation Wilkens et al. Most of these enzymes arise in isoforms exhibiting changes in specificity and ability to interact with different partners forming multi-protein complexes of functional relevance for the production of the polysaccharide Crofts et al.

The enzyme from different sources is allosterically regulated by metabolites that are critical intermediates of the central carbon energy metabolism operating in the respective organism Ballicora et al. Subunits S and L are homologous proteins, where the L polypeptide has emerged in different species via gene duplication followed by subfunctionalization Ballicora et al. In potato tuber, the S subunit retained the catalytic function, whereas the L subunit specialized in modulating the regulation of the former Ballicora et al.

The interaction between both subunits is determinant for the enzyme activity and regulatory responses Kavakli et al. Excluding the enzyme from monocot as wheat endosperm, the ADP-Glc PPase S subunit from leaves and other plant tissues has an N-terminal cysteine residue that is critical for redox regulation. This latter involves the formation of a disulfide bridge between the S subunits in the heterotetramer, mediated by the thioredoxin system Ballicora et al.

In addition, as it has been reviewed previously Ballicora et al. In the barley and maize forms, 3PGA modifies the relative affinity for substrates Plaxton and Preiss, ; Kleczkowski et al.

A substantial body of experimental evidence is giving support to the modulation of starch biosynthesis by the combined action of post-translational mechanisms, including redox modification, protein complex formation, and protein phosphorylation Lohrig et al.

In this context, proteomic information obtained in the last decade draws attention to ADP-Glc PPase as a putative target of protein kinases Lohrig et al. Specifically, a proteomic analysis of maize endosperm identified the phosphorylation of the small subunit of the enzyme Yu et al.

However, all these predictive results lack functional and developmental evidence of the tangible presence of the enzyme at a phosphorylated state in planta.

The latter is critical for the complete understanding of factors affecting plant productivity, thus limiting the design of better strategies for its improvement. Results suggest that phosphorylation of the enzyme involved in the limiting step of starch built-up would be functionally relevant for the yield of grains in grass crops. All other reagents were of the highest quality available. Seeds harvesting was as described previously Piattoni et al. Briefly, Triticum aestivum L.

Baguette 11 samples collected at 3, 6, 10, 14, 17, and 27 days post-anthesis DPA , and spikes were frozen immediately in liquid nitrogen. Castor Ricinus communis seeds were collected at 5, 10, 20, 25, 32, and 40 days post-pollination DPP.

Seeds whole protein extraction was made by triplicate using independent biological replicates as reported elsewhere Piattoni et al.

Frozen seeds of wheat or castor oil were ground to a fine powder in liquid nitrogen using a mortar and pestle. The mixture was incubated 20 min on ice with constant homogenization. The supernatant was recovered and used immediately for the different assays. Quantification of starch from seeds was as indicated in Piattoni et al.

The supernatant was discarded and the extraction repeated thrice to eliminate soluble sugars. To determine the contents of TAGs in seeds, we utilized protocols already described Folch et al. Plant tissue mg was ground to a fine powder in liquid nitrogen and lipids extracted with 0.

The extraction was performed in hermetically closed tubes incubated 2 h at room temperature with gentle mixing every 30 min. Samples were filtered on oil-free filter paper previously washed with the extraction solution, placed in previously weighed tubes, and vigorously mixed with 0. After weighing, the lipids dissolved in isopropanol served to quantify TAGs by an enzymatic colorimetric assay.

This method was based on the treatment of TAGs with lipase, then converting the produced glycerol into glycerol-3P by a specific kinase coupled to the reaction of glycerol-3P oxidase that produces H 2 O 2. The latter was quantified by employing a peroxidase generating a compound that absorbs at nm.

Assays initiated by the addition of Glc1P at a final concentration of 1. The complex formed with the released P i was measured at nm. Total protein quantification was determined by the method of Bradford Bradford, using BSA as a standard. The acrylamide-pendant Phos-tag ligand provides a molecule with a functional affinity to interact with phosphate groups, after which produce electrophoretic mobility-shifts of phosphorylated proteins Kinoshita et al.

The phosphorylation reaction was carried out as described below except for the use of non-radioactive ATP and modifying the reaction time to 2 h with the addition of the recombinant kinase and the corresponding amount of buffer according to the final volume change every 30 min. After the electrophoretic run 30 mA per gel , gels were washed twice in methanol-free Tris-Gly transfer buffer and 1 mM EDTA, for 10 min each time with gentle agitation, followed other 10 min in methanol-free transfer buffer without EDTA.

It followed electrotransference of the proteins and immunodetection with specific antibodies. Total proteins 1. The increase of pH in three steps was then employed to elute the adsorbed. The protein purification was performed from three independent biological replicates to enable an assessment of significance. Protein de-phosphorylation experiments followed a protocol described elsewhere Bustos and Iglesias, Then, samples were analyzed by electrophoresis followed by western blotting and immunodetection.

The Mdo SOS2 gene was synthesized de novo with a sequence and codon usage optimized for its expression in E. The TD mutation [required for activity Guo, ] was introduced by quick-change mutagenesis and then subcloned into a pET28b vector to produce a protein with an N-term His-tag. Cells were induced with 0. After centrifuging twice at 30, x g for 10 min, the supernatant crude extract was loaded onto a 1 ml HisTrap column GE Healthcare previously equilibrated with Buffer H.

The three I—III specific phosphorylation conditions used for assay crude extracts from seeds or purified recombinant protein kinase were as follows.

Worthy of mention is that the critical difference between these protein kinases is related to requirement or not of calcium. All phosphorylation assays were performed from three independent technical replicates to enable an assessment of significance. Seeds of grasses are relevant components within the world crops providing food and energy, with wheat being one of the principal edible grains.

From this, it is clear the relevance of starch as a major staple. Another primary product of agriculture is TAGs accumulated in seeds of oleaginous plants. To better understand starch biosynthesis and its regulation in seeds, we explored about contents of the polysaccharide and TAGs, as well as ADP-Glc PPase activity and phosphorylation profiles along with the development of Triticum aestivum wheat seeds Table 1 and Figure 1.

For comparison, we performed similar studies with seeds of the oleaginous Ricinus communis castor bean Supplemental Table 1 and Supplemental Figure 1. These profiles are in agreement with values reported in previous studies on developing wheat endosperm Briarty et al. Instead, castor bean seeds exhibited a progressive increase in the amount of starch during cell proliferation 0—20 DPP , but an abrupt decrease during accumulation of reserves 20—40 DPP and maturation plus desiccation 30—50 DPP.

DPA, days post-anthesis.