Trypanosomatid phosphoglycerate mutases have multiple conformational and oligomeric states

Blackburn, Elizabeth A.; Fuad, Fazia Adyani Ahmad; Morgan, Hugh P.; Nowicki, Matthew W.; Wear, Martin A.; Michels, Paul A.M.; Fothergill‐Gilmore, Linda A.; Walkinshaw, Malcolm D.

doi:10.1016/j.bbrc.2014.06.113

Cited by 9 publications

(7 citation statements)

References 24 publications

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“…Analysis of amino acid sequences shows that iPGMs occur in nature in two structurally different families. iPGMs found in plants and trypanosomatids belong to family 1 and the rest belong to family 2 . The metal dependence of these two classes is also different.…”

Section: Introductionmentioning

confidence: 98%

Complete catalytic cycle of cofactor‐independent phosphoglycerate mutase involves a spring‐loaded mechanism

et al. 2015

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Cofactor-independent phosphoglycerate mutase (iPGM), an important enzyme in glycolysis and gluconeogenesis, catalyses the isomerization of 2-and 3-phosphoglycerates by an Mn 2+ -dependent phospho-transfer mechanism via a phospho-enzyme intermediate. Crystal structures of bi-domain iPGM from Staphylococcus aureus, together with substrate-bound forms, have revealed a new conformation of the enzyme, representing an intermediate state of domain movement. The substrate-binding site and the catalytic site are present in two distinct domains in the intermediate form.X-ray crystallography complemented by simulated dynamics has enabled delineation of the complete catalytic cycle, which includes binding of the substrate, followed by its positioning into the catalytic site, phospho-transfer and finally product release. The present work describes a novel mechanism of domain movement controlled by a hydrophobic patch that is exposed on domain closure and acts like a spring to keep the protein in open conformation. Domain closing occurs after substrate binding, and is essential for phospho-transfer, whereas the open conformation is a prerequisite for efficient substrate binding and product dissociation. A new model of catalysis has been proposed by correlating the hinge-bending motion with the phospho-transfer mechanism.

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Section: Introductionmentioning

confidence: 98%

Complete catalytic cycle of cofactor‐independent phosphoglycerate mutase involves a spring‐loaded mechanism

et al. 2015

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“…Second, sequence alignments clearly indicated that S82 of At iPGAM2 corresponds to S62 of Sa iPGAM and Gs iPGAM, S74 of Tb iPGAM and S75 of Lm iPGAM (Blackburn et al. , 2014), which have been demonstrated to undergo transient phosphorylation as part of the catalytic reaction mechanism. Third, it was apparent that residues known to be involved in non‐plant iPGAM catalysis (e.g., see Nowicki et al.…”

Section: Resultsmentioning

confidence: 99%

“…Furthermore, iPGAM proteins can be classified into two families according to their amino acid sequence and cation dependence: family 1 is composed of enzymes found in Trypanosomatidae and plants and use Mg 2+ /Zn 2+ , while family 2 members can be found in prokaryotes and require Mn 2+ to be active (Blackburn et al. , 2014; Fuad et al. , 2011).…”

Section: Introductionmentioning

confidence: 99%

Arabidopsis thaliana 2,3‐bisphosphoglycerate‐independent phosphoglycerate mutase 2 activity requires serine 82 phosphorylation

et al. 2021

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SUMMARY Phosphoglycerate mutases (PGAMs) catalyse the reversible isomerisation of 3‐phosphoglycerate and 2‐phosphoglycerate, a step of glycolysis. PGAMs can be sub‐divided into 2,3‐bisphosphoglycerate‐dependent (dPGAM) and ‐independent (iPGAM) enzymes. In plants, phosphoglycerate isomerisation is carried out by cytosolic iPGAM. Despite its crucial role in catabolism, little is known about post‐translational modifications of plant iPGAM. In Arabidopsis thaliana, phosphoproteomics analyses have previously identified an iPGAM phosphopeptide where serine 82 is phosphorylated. Here, we show that this phosphopeptide is less abundant in dark‐adapted compared to illuminated Arabidopsis leaves. In silico comparison of iPGAM protein sequences and 3D structural modelling of AtiPGAM2 based on non‐plant iPGAM enzymes suggest a role for phosphorylated serine in the catalytic reaction mechanism. This is confirmed by the activity (or the lack thereof) of mutated recombinant Arabidopsis iPGAM2 forms, affected in different steps of the reaction mechanism. We thus propose that the occurrence of the S82‐phosphopeptide reflects iPGAM2 steady‐state catalysis. Based on this assumption, the metabolic consequences of a higher iPGAM activity in illuminated versus darkened leaves are discussed.

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“…In the closed form the enzyme (as in Fig. S1B) catalyses 2/3PGA isomerization resulting in product release, and product release causes opening of the enzyme and return to the open conformation (Blackburn et al 2014).…”

Section: Resultsmentioning

confidence: 99%

Molecular, biochemical characterization and assessment of immunogenic potential of cofactor-independent phosphoglycerate mutase againstLeishmania donovani: a step towards exploring novel vaccine candidate

et al. 2017

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Despite immense efforts, vaccine against visceral leishmaniasis has yet not been developed. Earlier our proteomic study revealed a novel protein, cofactor-independent phoshoglycerate mutase (LdiPGAM), an important enzyme in glucose metabolism, in T helper cells type 1 (Th1) stimulatory region of soluble Leishmania donovani antigen. In this study, LdiPGAM was biochemically and molecularly characterized and evaluated for its immunogenicity and prophylactic efficacy against L. donovani. Immunogenicity of recombinant LdiPGAM (rLdiPGAM) was initially assessed in naïve hamsters immunized with it by analysing mRNA expression of inducible nitric oxide (NO) synthase (iNOS) and other Th1/T helper cells type 2 cytokines, which revealed an upregulation of Th1 cytokines along with iNOS. Immunogenicity of rLdiPGAM was further evaluated in lymphocytes of treated Leishmania-infected hamsters and peripheral blood mononuclear cells of Leishmania patients in clinical remission by various parameters, viz. lymphoproliferation assay and NO production (hamsters and patients) and levels of various cytokines (patients). rLdiPGAM induced remarkable Lymphoproliferative response and NO production in treated Leishmania-infected hamsters as well as in patients and increase in interferon gamma (IFN-γ), interleukin-12 (IL-12p40) responses in Leishmania patients in clinical remission. Vaccination with rLdiPGAM exerted considerable prophylactic efficacy (73%) supported by increase in mRNA expression of iNOS, IFN-γ and IL-12p40 with decrease in transforming growth factor beta and interleukin-10. Above results indicate the importance of rLdiPGAM protein as a potential vaccine candidate against visceral leishmaniasis.

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Trypanosomatid phosphoglycerate mutases have multiple conformational and oligomeric states

Cited by 9 publications

References 24 publications

Complete catalytic cycle of cofactor‐independent phosphoglycerate mutase involves a spring‐loaded mechanism

Complete catalytic cycle of cofactor‐independent phosphoglycerate mutase involves a spring‐loaded mechanism

Arabidopsis thaliana 2,3‐bisphosphoglycerate‐independent phosphoglycerate mutase 2 activity requires serine 82 phosphorylation

Molecular, biochemical characterization and assessment of immunogenic potential of cofactor-independent phosphoglycerate mutase againstLeishmania donovani: a step towards exploring novel vaccine candidate

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