Mycobacterial OtsA Structures Unveil Substrate Preference Mechanism and Allosteric Regulation by 2-Oxoglutarate and 2-Phosphoglycerate

Mendes, V.; Acebrón-García-de-Eulate, Marta; Verma, Nandini; Błaszczyk, M.; Dias, M.V.B.; Blundell, Tom L.

doi:10.1128/mbio.02272-19

Cited by 10 publications

(11 citation statements)

References 62 publications

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“…Trehalose-6-phosphate synthase (TPS) catalyzes the synthesis of trehalose, which is primarily synthesized in insects’ fat bodies via the TPS/trehalose-6-phosphate phosphatase pathway [ 27 , 28 , 29 ]. Compounds such as trehalose play a crucial role in the negative feedback regulation of TPS activity [ 30 , 31 ]. Evidence indicates that the capacity of TPS accumulation varies with temperature induction, as noted in the TPS levels of Drosophila melanogaster [ 32 ], Delia antiqua [ 33 ], Maruca vitrata [ 34 ], and Megaphorura arctica [ 35 ].…”

Section: Introductionmentioning

confidence: 99%

Fruit Fly in a Challenging Environment: Impact of Short-Term Temperature Stress on the Survival, Development, Reproduction, and Trehalose Metabolism of Bactrocera dorsalis (Diptera: Tephritidae)

Zhao

Zhou

et al. 2022

Insects

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An understanding of physiological damage and population development caused by uncomfortable temperature plays an important role in pest control. In order to clarify the adaptability of different temperatures and physiological response mechanism of B. dorsalis, we focused on the adaptation ability of this pest to environmental stress from physiological and ecological viewpoints. In this study, we explored the relationship between population parameters and glucose, glycogen, trehalose, and trehalose-6-phosphate synthase responses to high and low temperatures. Compared with the control group, temperature stress delayed the development duration of all stages, and the survival rates and longevity decreased gradually as temperature decreased to 0 °C and increased to 36 °C. Furthermore, with low temperature decrease from 10 °C to 0 °C, the average fecundity per female increased at 10 °C but decreased later. Reproduction of the species was negatively affected during high-temperature stresses, reaching the lowest value at 36 °C. In addition to significantly affecting biological characteristics, temperature stress influenced physiological changes of B. dorsalis in cold and heat tolerance. When temperature deviated significantly from the norm, the levels of substances associated with temperature resistance were altered: glucose, trehalose, and TPS levels increased, but glycogen levels decreased. These results suggest that temperature stresses exert a detrimental effect on the populations’ survival, but the metabolism of trehalose and glycogen may enhance the pest’s temperature resistance.

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Section: Introductionmentioning

confidence: 99%

Fruit Fly in a Challenging Environment: Impact of Short-Term Temperature Stress on the Survival, Development, Reproduction, and Trehalose Metabolism of Bactrocera dorsalis (Diptera: Tephritidae)

Zhao

Zhou

et al. 2022

Insects

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“…Both structures are superimposed on the C-terminal domain to highlight the relative movement of domains. (D) On the right side, the superposition of M. thermoresistibile OtsA as reference (PDB 5K44) 440 with superimposed substrates ADP and glucose 6-phosphate (PDB 5JIO, ligands in black), 440 ADP-glucose (PDB 5K41, ligand in green), 440 and GDP-glucose (PDB 5K42, ligand semitransparent). 440 Note the GDP-glucose nucleotide ring in a displaced position with respect the preferred substrate ADP-glucose.…”

Section: The Gt-b Fold Enzymesmentioning

confidence: 99%

“…(D) On the right side, the superposition of M. thermoresistibile OtsA as reference (PDB 5K44) 440 with superimposed substrates ADP and glucose 6-phosphate (PDB 5JIO, ligands in black), 440 ADP-glucose (PDB 5K41, ligand in green), 440 and GDP-glucose (PDB 5K42, ligand semitransparent). 440 Note the GDP-glucose nucleotide ring in a displaced position with respect the preferred substrate ADP-glucose. The left panel shows the reference structure MtOtsA in complex with trehalose 6-phosphate (PDB 5K44, ligand in pink) 440 in the superposition with ADP (PDB 5JIO).…”

Section: The Gt-b Fold Enzymesmentioning

confidence: 99%

“…In addition, OtsA from M. thermoresistibile complexes with ADP-glucose or GDP-glucose highlight subtle differences in the binding site that explains the preference for ADP-glucose. 440 Interestingly, three single-point mutations allow the conversion of the donor substrate preference to UDP-glucose.…”

Section: Trehalose 6-phosphate Synthase Otsa and Trehalose Synthasementioning

confidence: 99%

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Architecture, Function, Regulation, and Evolution of α-Glucans Metabolic Enzymes in Prokaryotes

Cifuente,

Colleoni,

Kalscheuer

et al. 2024

Chem. Rev.

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Bacteria have acquired sophisticated mechanisms for assembling and disassembling polysaccharides of different chemistry. α-D-Glucose homopolysaccharides, so-called α-glucans, are the most widespread polymers in nature being key components of microorganisms. Glycogen functions as an intracellular energy storage while some bacteria also produce extracellular assorted α-glucans. The classical bacterial glycogen metabolic pathway comprises the action of ADP-glucose pyrophosphorylase and glycogen synthase, whereas extracellular α-glucans are mostly related to peripheral enzymes dependent on sucrose. An alternative pathway of glycogen biosynthesis, operating via a maltose 1phosphate polymerizing enzyme, displays an essential wiring with the trehalose metabolism to interconvert disaccharides into polysaccharides. Furthermore, some bacteria show a connection of intracellular glycogen metabolism with the genesis of extracellular capsular α-glucans, revealing a relationship between the storage and structural function of these compounds. Altogether, the current picture shows that bacteria have evolved an intricate α-glucan metabolism that ultimately relies on the evolution of a specific enzymatic machinery. The structural landscape of these enzymes exposes a limited number of core catalytic folds handling many different chemical reactions. In this Review, we present a rationale to explain how the chemical diversity of α-glucans emerged from these systems, highlighting the underlying structural evolution of the enzymes driving α-glucan bacterial metabolism.

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“…OtsA is a member of family 20 glycosyltransferase (GT20) that catalyzes a coupling reaction between nucleotidyl diphosphate (NDP)-glucose and glucose 6-phosphate to give α,α-1,1-trehalose-6 phosphate as a product. ,, X-ray crystal structures of OtsA in complex with uridine diphosphate (UDP) and a transition state mimicry, validoxylamine A 7′-phosphate (VDO), as well as results from a kinetic isotope effect study suggest the involvement of an oxocarbenium ion-like transition state in OtsA catalysis. , Moreover, this mechanism involves hydrogen bonding between the leaving NDP group and the acceptor nucleophile, which allows a front-face nucleophilic attack to form a glycosidic bond with retained stereochemistry. The leaving group phosphate also acts as a general base that deprotonates the acceptor nucleophile …”

Section: Introductionmentioning

confidence: 99%

Catalytic Mechanism of Nonglycosidic C–N Bond Formation by the Pseudoglycosyltransferase Enzyme VldE

Tsunoda,

Abuelizz,

Samadi

et al. 2023

ACS Catal.

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The pseudoglycosyltransferase (PsGT) enzyme VldE is a homologue of the retaining glycosyltransferase (GT) trehalose 6-phosphate synthase (OtsA) that catalyzes a coupling reaction between two pseudosugar units, GDP-valienol and validamine 7-phosphate, to give a product with α,α-N-pseudoglycosidic linkage. Despite its biological importance and unique catalytic function, the molecular bases for its substrate specificity and reaction mechanism are still obscure. Here, we report a comparative mechanistic study of VldE and OtsA using various engineered chimeric proteins and point mutants of the enzymes, X-ray crystallography, docking studies, and kinetic isotope effects. We found that the distinct substrate specificities between VldE and OtsA are most likely due to topological differences within the hot spot amino acid regions of their N-terminal domains. We also found that the Asp158 and His182 residues, which are in the active site, play a significant role in the PsGT function of VldE. They do not seem to be directly involved in the catalysis but may be important for substrate recognition or contribute to the overall architecture of the active site pocket. Moreover, results of the kinetic isotope effect experiments suggest that VldE catalyzes C–N bond formation between GDP-valienol and validamine 7-phosphate via an SN i-like mechanism. The study provides new insights into the substrate specificity and catalytic mechanism of a member of the growing family of PsGT enzymes, which may be used as a basis for developing new PsGTs from GTs.

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Mycobacterial OtsA Structures Unveil Substrate Preference Mechanism and Allosteric Regulation by 2-Oxoglutarate and 2-Phosphoglycerate

Cited by 10 publications

References 62 publications

Fruit Fly in a Challenging Environment: Impact of Short-Term Temperature Stress on the Survival, Development, Reproduction, and Trehalose Metabolism of Bactrocera dorsalis (Diptera: Tephritidae)

Fruit Fly in a Challenging Environment: Impact of Short-Term Temperature Stress on the Survival, Development, Reproduction, and Trehalose Metabolism of Bactrocera dorsalis (Diptera: Tephritidae)

Architecture, Function, Regulation, and Evolution of α-Glucans Metabolic Enzymes in Prokaryotes

Catalytic Mechanism of Nonglycosidic C–N Bond Formation by the Pseudoglycosyltransferase Enzyme VldE

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