Stereospecific chemoenzymatic synthesis of the deuterated analogues of 2-phosphoglycerate and structural analysis by means of NMR spectroscopy

Vialletelle, V.; Rabiller, Claude; Heisler, A.; Levayer, F.

doi:10.1016/s0957-4166(00)80497-8

Cited by 2 publications

(1 citation statement)

References 6 publications

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“…18 O]PEP disodium salt was prepared using slight modifications to literature-reported procedures (49,50). The resulting material was 66% enriched in 18 O isotope at the C-2 position as determined by both 31 P NMR spectroscopy and mass spectrometry.…”

Section: O-labeling Experiment-[2-mentioning

confidence: 99%

Structural and Mechanistic Analysis of Sialic Acid Synthase NeuB from Neisseria meningitidis in Complex with Mn2+, Phosphoenolpyruvate, and N-Acetylmannosaminitol

Gunawan

Simard

Gilbert³

et al. 2005

Journal of Biological Chemistry

137

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In Neisseria meningitidis and related bacterial pathogens, sialic acids play critical roles in mammalian cell immunity evasion and are synthesized by a conserved enzymatic pathway that includes sialic acid synthase (NeuB, SiaC, or SynC). NeuB catalyzes the condensation of phosphoenolpyruvate (PEP) and N-acetylmannosamine, directly forming N-acetylneuraminic acid (or sialic acid). In this paper we report the development of a coupled assay to monitor NeuB reaction kinetics and an 18 O-labeling study that demonstrates the synthase operates via a COO bond cleavage mechanism. We also report the first structure of a sialic acid synthase, that of NeuB, revealing a unique domain-swapped homodimer architecture consisting of a (␤/␣) 8 barrel (TIM barrel)-type fold at the N-terminal end and a domain with high sequence identity and structural similarity to the ice binding type III antifreeze proteins at the C-terminal end of the enzyme. We have determined the structures of NeuB in the malate-bound form and with bound PEP and the substrate analog N-acetylmannosaminitol to 1.9 and 2.2 Å resolution, respectively. Typical of other TIM barrel proteins, the active site of NeuB is located in a cavity at the C-terminal end of the barrel; however, the positioning of the swapped antifreeze-like domain from the adjacent monomer provides key residues for hydrogen bonding with substrates in the active site of NeuB, a structural feature that leads to distinct modes of substrate binding from other PEP-utilizing enzymes that lack an analogous antifreeze-like domain. Our observation of a direct interaction between a highly ordered manganese and the N-acetylmannosaminitol in the NeuB active site also suggests an essential role for the ion as an electrophilic catalyst that activates the Nacetylmannosamine carbonyl to the addition of PEP.

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Section: O-labeling Experiment-[2-mentioning

confidence: 99%

Structural and Mechanistic Analysis of Sialic Acid Synthase NeuB from Neisseria meningitidis in Complex with Mn2+, Phosphoenolpyruvate, and N-Acetylmannosaminitol

Gunawan

Simard

Gilbert³

et al. 2005

Journal of Biological Chemistry

137

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Effets des sels de terres rares sur quelques phosphates dérivés des metabolites de la glycolyse

Rabiller¹,

Eymard²,

Mesbahi³

1991

Can. J. Chem.

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R e p le 13 dtcembre 1990 CLAUDE RABILLER, PHIL~PPE EYMARD et MAHMOUD MESBAHI. Can. J. Chem. 69, 1281Chem. 69, (1991.Les dtrivts phosphatts du glucose et du fructose sont des intermtdiaires importants dans la fermentation des sucres. L'ttude du devenir de leurs protons (ou deuttrons) par spectroscopie RMN doit fournir des informations sur les transferts de protons (ou deuterons) susceptibles d'intervenir au cours de cette bio-transformation. Dans cette note, nous dkcrivons I'influence de la complexation de ces mttabolites avec le chlorure de prastodyme ainsi que celle du pH sur leurs dtplacements chimiques protoniques. On dtcrit par ailleurs les conditions dans lesquelles tous les protons des formes cr et P de ces sucres cycliques sont individualists. On a trouvt que le glucose-1-phosphate prtsente les meilleures aptitudes pour ce type d'etude. De plus, la comparaison du comportement de ces composts avec celui de l'acide 2-phosphoglyctrique permet de prtciser la conformation de ce dernier.Mots cle's : dtrivCs phosphatts des sucres, RMN, effets des terres rares. CLAUDE RABILLER, PHILIPPE EYMARD, and MAHMOUD MESBAHI. Can. J. Chem. 69, 1281Chem. 69, (1991.Phosphate derivatives of glucose and fructose are well-known intermediates in sugar fermentation. Proton and deuterium NMR spectroscopy studies of the fermentation process provide information about proton (deuteron) transfers that are likely to occur in such biotransformations. In this paper we describe the influence of complexation of those metabolites with praseodymium trichloride and of pH on their proton NMR chemical shifts. We determine the conditions under which all the protons of both the cr and p forms of these cyclic sugars are separated. Glucose-1 phosphate was found to be the most suitable derivative for this type of study. Furthermore, comparing the behaviour of these compounds with 2-phosphoglyceric acid allows us to specify the exact conformation of the latter.

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Stereospecific chemoenzymatic synthesis of the deuterated analogues of 2-phosphoglycerate and structural analysis by means of NMR spectroscopy

Cited by 2 publications

References 6 publications

Structural and Mechanistic Analysis of Sialic Acid Synthase NeuB from Neisseria meningitidis in Complex with Mn2+, Phosphoenolpyruvate, and N-Acetylmannosaminitol

Structural and Mechanistic Analysis of Sialic Acid Synthase NeuB from Neisseria meningitidis in Complex with Mn2+, Phosphoenolpyruvate, and N-Acetylmannosaminitol

Effets des sels de terres rares sur quelques phosphates dérivés des metabolites de la glycolyse

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