The Structure of Sucrose Synthase-1 from Arabidopsis thaliana and Its Functional Implications

Zheng, Yi; Anderson, Spencer; Zhang, Yanfeng; Garavito, R. Michael

doi:10.1074/jbc.m111.275974

Cited by 90 publications

(108 citation statements)

References 58 publications

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“…A similar result was observed in the complexes between ADP-Glc and EcGS (24), and between UDP-Glc and Arabidopsis thaliana sucrose synthase, a member of the retaining GT4 family (19). In those cases, however, the respective diphosphonucleoside moieties of the donor molecule were present in the active sites of the enzymes, although not covalently bound to the not precisely identified glucose derivative.…”

Section: Discussionsupporting

confidence: 70%

“…Further along these lines, the discrete existence of a short-lived ion pair intermediate in the reaction pathway (Fig. 1C), which would result from the asynchronous cleavage of the bond between the anomeric carbon and leaving group and the formation of the donor-acceptor bond, has been proposed (4,18,19). According to IUPAC recommendations, this stepwise mechanism would be represented by D N *A N , where the asterisk indicates the formation of a short-lived intermediate.…”

Section: Introductionmentioning

confidence: 88%

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Lyase activity of glycogen synthase: Is an elimination/addition mechanism a possible reaction pathway for retaining glycosyltransferases?

et al. 2012

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Despite the biological relevance of glycosyltrasferases (GTs) and the many efforts devoted to this subject, the catalytic mechanism through which a subclass of this large family of enzymes, namely those that operate with net retention of the anomeric configuration, has not been fully established. Here, we show that in the absence of an acceptor, an archetypal retaining GT such as Pyrococcus abyssi glycogen synthase (PaGS) reacts with its glucosyl donor substrate, uridine 5'‐diphosphoglucose (UDP‐Glc), to produce the scission of the covalent bond between the terminal phosphate oxygen of UDP and the sugar ring. X‐ray diffraction analysis of the PaGS/UDP‐Glc complex shows no electronic density attributable to the UDP moiety, but establishes the presence in the active site of the enzyme of a glucose‐like derivative that lacks the exocyclic oxygen attached to the anomeric carbon. Chemical derivatization followed by gas chromatography/mass spectrometry of the isolated glucose‐like species allowed us to identify the molecule found in the catalytic site of PaGS as 1,5‐anhydro‐D‐arabino‐hex‐1‐enitol (AA) or its tautomeric form, 1,5‐anhydro‐D‐fructose. These findings are consistent with a stepwise SNi‐like mechanism as the modus operandi of retaining GTs, a mechanism that involves the discrete existence of an oxocarbenium intermediate. Even in the absence of a glucosyl acceptor, glycogen synthase (GS) promotes the formation of the cationic intermediate, which, by eliminating the proton of the adjacent C2 carbon atom, yields AA. Alternatively, these observations could be interpreted assuming that AA is a true intermediate in the reaction pathway of GS and that this enzyme operates through an elimination/addition mechanism. © 2012 IUBMB Life, 64(7): 649–658, 2012.

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Section: Discussionsupporting

confidence: 70%

Section: Introductionmentioning

confidence: 88%

Lyase activity of glycogen synthase: Is an elimination/addition mechanism a possible reaction pathway for retaining glycosyltransferases?

et al. 2012

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“…Therefore, understanding the catalysis and the regulation of sucrose synthase is of great significance. Recently, Zheng et al (23) reported the crystal structure of the Arabidopsis thaliana sucrose synthase in complex with UDP and fructose in a closed conformation. This enzyme is a homotetramer composed of four identical subunits of ϳ90 kDa and belongs to group 4 of the GT-B retaining glycosyltransferase family (http://www.cazy.org/GlycosylTransferases.html) (24).…”

mentioning

confidence: 99%

“…This enzyme is a homotetramer composed of four identical subunits of ϳ90 kDa and belongs to group 4 of the GT-B retaining glycosyltransferase family (http://www.cazy.org/GlycosylTransferases.html) (24). An S N i-like reaction mechanism has been proposed for this enzyme family (23)(24)(25).…”

mentioning

confidence: 99%

The Crystal Structure of Nitrosomonas europaea Sucrose Synthase Reveals Critical Conformational Changes and Insights into Sucrose Metabolism in Prokaryotes

Diez

Figueroa

et al. 2015

J Bacteriol

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In this paper we report the first crystal structure of a prokaryotic sucrose synthase from the nonphotosynthetic bacterium Nitrosomonas europaea. The obtained structure was in an open form, whereas the only other available structure, from the plant Arabidopsis thaliana, was in a closed conformation. Comparative structural analysis revealed a "hinge-latch" combination, which is critical to transition between the open and closed forms of the enzyme. The N. europaea sucrose synthase shares the same fold as the GT-B family of the retaining glycosyltransferases. In addition, a triad of conserved homologous catalytic residues in the family was shown to be functionally critical in the N. europaea sucrose synthase (Arg567, Lys572, and Glu663). This implies that sucrose synthase shares not only a common origin with the GT-B family but also a similar catalytic mechanism. The enzyme preferred transferring glucose from ADP-glucose rather than UDP-glucose like the eukaryotic counterparts. This predicts that these prokaryotic organisms have a different sucrose metabolic scenario from plants. Nucleotide preference determines where the glucose moiety is targeted after sucrose is degraded. IMPORTANCEWe obtained biochemical and structural evidence of sucrose metabolism in nonphotosynthetic bacteria. Until now, only sucrose synthases from photosynthetic organisms have been characterized. Here, we provide the crystal structure of the sucrose synthase from the chemolithoautotroph N. europaea. The structure supported that the enzyme functions with an open/close induced fit mechanism. The enzyme prefers as the substrate adenine-based nucleotides rather than uridine-based like the eukaryotic counterparts, implying a strong connection between sucrose and glycogen metabolism in these bacteria. Mutagenesis data showed that the catalytic mechanism must be conserved not only in sucrose synthases but also in all other retaining GT-B glycosyltransferases. In plants, sucrose is a major photosynthetic product and plays a key role not only for carbon partition but also in sugar sensing, development, and regulation of gene expression (1-3). It was first thought that sucrose metabolism was a characteristic of plants, but it was later found in other oxygenic photosynthetic organisms (4, 5). In the last decade, Salerno and coworkers demonstrated the importance of sucrose for carbon and nitrogen fixation in filamentous cyanobacteria (6, 7). More recently, genomic and phylogenetic analyses revealed the existence of sucrose-related genes in nonphotosynthetic prokaryotes such as proteobacteria, firmicutes, and planctomycetes (4, 5, 8). It has been suggested that these organisms acquired the genes of sucrose metabolism by horizontal gene transfer (4,5,8). However, analysis of the enzymes encoded by such genes is currently lacking.Nitrosomonas europaea is a chemolithoautotrophic bacterium that obtains energy by oxidizing ammonia to hydroxylamine and nitrite in the presence of oxygen (9). It is a member of the betaproteobacteria group with a putati...

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“…Cassava leaf SuS plays an important role in the distribution of the photosynthesized carbohydrate (Zheng et al, 2011), thereby affects the cassava tuber starch accumulation. The cassava leaf SuS synthesis and decomposing direction activities were determined during the growth period (Figure 1 and 2).…”

Section: The Time Course Of Cassava Leaf Sus Synthesis Direction and mentioning

confidence: 99%

Time Course of Starch Biosynthesis Enzymes Activity and Root Tuber Starch of Four Cassava Cultivars

Huang¹,

Luo²,

Huang³

et al. 2017

JAS

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To understand the accumulation rule of cassava root tuber starch, the amylose, amylopectin and total starch content of fresh root tuber, the enzyme activities of sucrose synthase (SuS, EC 2.4.1.13) and sucrose phosphate synthase (SPS, EC 2.4.1.14) of leaves, the enzyme activities of ADP-glucose pyrophosphorylase (AGPase, EC 2.7.7.27), soluble starch synthase (SSS, EC 2.4.1.21) and starch branching enzyme (SBE, EC. 2.4.1.18) of tubers were assessed by using cultivars SC201, SC205, GR891 and GR911 leaves and root tubers during their growth period, respectively. The results as follows: the enzyme activity of leaf SPS and the synthesis direction of SuS showed the highest at August, 2010, however the enzyme activity of the decomposing direction of SuS formed parabolic curve; the enzyme activity of tuber AGPase showed an increased then decreased single peak curve, the enzyme activity of tuber SSS oscillating decreased, while the enzyme activity of SBE was relatively stable; the amylose, amylopectin and total starch contents of fresh cassava tuber were all gradually increased along with the growth period. This research would enrich our knowledge of the time course of amylose, amylopectin and total starch contents of fresh cassava tuber, and above related enzyme activities.

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The Structure of Sucrose Synthase-1 from Arabidopsis thaliana and Its Functional Implications

Cited by 90 publications

References 58 publications

Lyase activity of glycogen synthase: Is an elimination/addition mechanism a possible reaction pathway for retaining glycosyltransferases?

Lyase activity of glycogen synthase: Is an elimination/addition mechanism a possible reaction pathway for retaining glycosyltransferases?

The Crystal Structure of Nitrosomonas europaea Sucrose Synthase Reveals Critical Conformational Changes and Insights into Sucrose Metabolism in Prokaryotes

Time Course of Starch Biosynthesis Enzymes Activity and Root Tuber Starch of Four Cassava Cultivars

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