Sucrose and starch are end products of two segregated gluconeogenic pathways, and their production takes place in the cytosol and chloroplast of green leaves, respectively. According to this view, the plastidial ADP⅐glucose (ADPG) pyrophosphorylase (AGP) is the sole enzyme catalyzing the synthesis of the starch precursor molecule ADPG. However, a growing body of evidences indicates that starch formation involves the import of cytosolic ADPG to the chloroplast. This evidence is consistent with the idea that synthesis of the ADPG linked to starch biosynthesis takes place in the cytosol by means of sucrose synthase, whereas AGP channels the glucose units derived from the starch breakdown. To test this hypothesis, we first investigated the subcellular localization of ADPG. Toward this end, we constructed transgenic potato plants that expressed the ADPG-cleaving adenosine diphosphate sugar pyrophosphatase (ASPP) from Escherichia coli either in the chloroplast or in the cytosol. Source leaves from plants expressing ASPP in the chloroplast exhibited reduced starch and normal ADPG content as compared with control plants. Most importantly however, leaves from plants expressing ASPP in the cytosol showed a large reduction of the levels of both ADPG and starch, whereas hexose phosphates increased as compared with control plants. No pleiotropic changes in photosynthetic parameters and maximum catalytic activities of enzymes closely linked to starch and sucrose metabolism could be detected in the leaves expressing ASPP in the cytosol. The overall results show that, essentially similar to cereal endosperms, most of the ADPG linked to starch biosynthesis in source leaves occurs in the cytosol.
A distinct phosphodiesterasic activity (EC 3.1.4) was found in both mono-and dicotyledonous plants that catalyzes the hydrolytic breakdown of ADPglucose (ADPG) to produce equimolar amounts of glucose-1-phosphate and AMP. The enzyme responsible for this activity, referred to as ADPG pyrophosphatase (AGPPase), was purified over 1,100-fold from barley leaves and subjected to biochemical characterization. The calculated K eq (modified equilibrium constant) value for the ADPG hydrolytic reaction at pH 7.0 and 25°C is 110, and its standard-state free-energy change value (⌬G) is ؊2.9 kcal͞mol (1 kcal ؍ 4.18 kJ). Kinetic analyses showed that, although AGPPase can hydrolyze several low-molecular weight phosphodiester bond-containing compounds, ADPG proved to be the best substrate (K m ؍ 0.5 mM). Pi and phosphorylated compounds such as 3-phosphoglycerate, PP i, ATP, ADP, NADP ؉ , and AMP are inhibitors of AGPPase. Subcellular localization studies revealed that AGPPase is localized exclusively in the plastidial compartment of cultured cells of sycamore (Acer pseudoplatanus L.), whereas it occurs both inside and outside the plastid in barley endosperm. In this paper, evidence is presented that shows that AGPPase, whose activity declines concomitantly with the accumulation of starch during development of sink organs, competes with starch synthase (ADPG:1,4-␣-D-glucan 4-␣-D-glucosyltransferase; EC 2.4.1.21) for ADPG, thus markedly blocking the starch biosynthesis.A lthough the pyrophosphorolytic reactions leading to the production of gluconeogenic intermediates such as ADPglucose (ADPG) and UDPglucose (UDPG) are readily reversible, they mainly proceed toward the direction of nucleotide sugar synthesis (1). On the other hand, plant enzymes that irreversibly cleave nucleotide sugars have been described that may effectively interrupt the flow of glycosyl moieties toward the biosynthesis of end products such as starch, cell wall polysaccharides, or sucrose (2). It is conceivable that in conjunction with ADPG pyrophosphorylase (AGPase; EC 2.7.7.27), UDPG pyrophosphorylase, sucrose synthase, and starch synthase, among others, these enzymes will participate in controlling the levels of nucleotide sugars engaged in starch formation. Among a few enzymes reported to date that can hydrolyze nucleotide sugars in plants, ADPG phosphorylase is shown to catalyze the phosphorolytic breakdown of ADPG (3). UDPG phosphorylase is known to split UDPG in the presence of P i , and its activities are greatly stimulated by some signal metabolites, indicating that it may play an important role in the control of photosynthate partitioning (4).Based on experimental grounds, Pozueta-Romero et al. (5) have proposed the operation of synthesis͞breakdown metabolic cycles controlling the rate of starch formation. According to this hypothesis, the balance between enzymatic activities catalyzing the synthesis of gluconeogenic intermediates and those activities catalyzing their breakdown can determine the net rate of starch synthesis. Because of the possibil...
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.