The structure of a <i>Trypanosoma cruzi</i> glucose‐6‐phosphate dehydrogenase reveals differences from the mammalian enzyme

Mercaldi, Gustavo Fernando; Dawson, Alice; Hunter, W.N.; Cordeiro, Artur T.

doi:10.1002/1873-3468.12276

Cited by 23 publications

(21 citation statements)

References 42 publications

(56 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Finally, we report the first 3D structural model of the G6PD domain of the G6PD::6PGL from G. lamblia , with important differences compared to the human G6PD enzyme. These structural dissimilarities with respect to human G6PD make the G6PD::6PGL from G. lamblia an ideal target for drug development, as the same approach has been used successfully in other parasites such as P. falciparum and Trypanosoma cruzy [ 14 , 26 , 41 ]. In P. falciparum , the 3D structure of PfG6PD‒6PGL compared to the human enzyme (G6PD) was reported, where a key difference in the substrate-binding site was shown that involves the replacement of Arg365 in human by Asp750 in PfG6PD.…”

Section: Resultsmentioning

confidence: 99%

Biochemical Characterization and Structural Modeling of Fused Glucose-6-Phosphate Dehydrogenase-Phosphogluconolactonase from Giardia lamblia

Morales-Luna

Serrano‐Posada

González‐Valdez

et al. 2018

IJMS

View full text Add to dashboard Cite

Glucose-6-phosphate dehydrogenase (G6PD) is the first enzyme in the pentose phosphate pathway and is highly relevant in the metabolism of Giardia lamblia. Previous reports suggested that the G6PD gene is fused with the 6-phosphogluconolactonase (6PGL) gene (6pgl). Therefore, in this work, we decided to characterize the fused G6PD-6PGL protein in Giardia lamblia. First, the gene of g6pd fused with the 6pgl gene (6gpd::6pgl) was isolated from trophozoites of Giardia lamblia and the corresponding G6PD::6PGL protein was overexpressed and purified in Escherichia coli. Then, we characterized the native oligomeric state of the G6PD::6PGL protein in solution and we found a catalytic dimer with an optimum pH of 8.75. Furthermore, we determined the steady-state kinetic parameters for the G6PD domain and measured the thermal stability of the protein in both the presence and absence of guanidine hydrochloride (Gdn-HCl) and observed that the G6PD::6PGL protein showed alterations in the stability, secondary structure, and tertiary structure in the presence of Gdn-HCl. Finally, computer modeling studies revealed unique structural and functional features, which clearly established the differences between G6PD::6PGL protein from G. lamblia and the human G6PD enzyme, proving that the model can be used for the design of new drugs with antigiardiasic activity. These results broaden the perspective for future studies of the function of the protein and its effect on the metabolism of this parasite as a potential pharmacological target.

show abstract

Section: Resultsmentioning

confidence: 99%

Biochemical Characterization and Structural Modeling of Fused Glucose-6-Phosphate Dehydrogenase-Phosphogluconolactonase from Giardia lamblia

Morales-Luna

Serrano‐Posada

González‐Valdez

et al. 2018

IJMS

View full text Add to dashboard Cite

show abstract

“…Mercaldi et al (52) reported that this G6PDH displays a key amino acid residue in the 2-2 domain, R72, which specifically interacts with the 2-phosphate group of NADPH (Fig. 6A).…”

Section: Resultsmentioning

confidence: 97%

“…One elegant example in this direction is the analysis of the (unique) G6PDH enzyme of the parasitic euglenoid Trypanosoma cruzi (51). Mercaldi et al (52) reported that this G6PDH displays a key amino acid residue in the β2-α2 domain, R72, which specifically interacts with the 2′-phosphate group of NADPH (Fig. 6A) .…”

Section: Resultsmentioning

confidence: 98%

See 1 more Smart Citation

Cofactor specificity of glucose-6-phosphate dehydrogenase isozymes inPseudomonas putidareveals a general principle underlying glycolytic strategies in bacteria

Volke

Olavarría

Nikel

2021

Preprint

View full text Add to dashboard Cite

Glucose-6-phosphate dehydrogenase (G6PDH) is widely distributed in nature and catalyzes the first committing step in the oxidative branch of the pentose phosphate (PP) pathway, feeding either the reductive PP or the Entner-Doudoroff pathway. Besides its role in central carbon metabolism, this dehydrogenase also provides reduced cofactors, thereby affecting redox balance. Although G6PDH is typically considered to display specificity towards nicotinamide adenine dinucleotide phosphate (NADP+), some variants accept nicotinamide NAD+ similarly (or even preferentially). Furthermore, the number of G6PDH isozymes encoded in bacterial genomes varies from none to more than four orthologues. On this background, we systematically analyzed the interplay of the three G6PDH isoforms of the soil bacterium Pseudomonas putida KT2440 from a genomic, genetic and biochemical perspective. P. putida represents an ideal model to tackle this endeavor, as its genome encodes numerous gene orthologues for most dehydrogenases in central carbon metabolism. We show that the three G6PDHs of strain KT2440 have different cofactor specificities, and that the isoforms encoded by zwfA and zwfB carry most of the activity, acting as metabolic ‘gatekeepers’ for carbon sources that enter at different nodes of the biochemical network. Moreover, we demonstrate how multiplication of G6PDH isoforms is a widespread strategy in bacteria, correlating with the presence of an incomplete Embden-Meyerhof-Parnas pathway. Multiplication of G6PDH isoforms in these species goes hand-in-hand with low NADP+ affinity at least in one G6PDH isozyme. We propose that gene duplication and relaxation in cofactor specificity is an evolutionary strategy towards balancing the relative production of NADPH and NADH.ImportanceProtein families have likely arisen during evolution by gene duplication and divergence followed by neo-functionalization. While this phenomenon is well documented for catabolic activities (typical of environmental bacteria that colonize highly polluted niches), the co-existence of multiple isozymes in central carbon catabolism remains relatively unexplored. We have adopted the metabolically-versatile soil bacterium Pseudomonas putida KT2440 as a model to interrogate the physiological and evolutionary significance of co-existing glucose-6-phosphate dehydrogenase (G6PDH) isozymes. Our results show that each of the three G6PDHs encoded in this bacterium display distinct biochemical properties, especially at the level of cofactor preference, impacting bacterial physiology in a carbon source-dependent fashion. Furthermore, the presence of multiple G6PDHs differing in NAD+- or NADP+-specificity in bacterial species strongly correlates with their predominant metabolic lifestyle. Our findings support the notion that multiplication of genes encoding cofactor-dependent dehydrogenases is a general evolutionary strategy towards achieving redox balance according to the growth conditions.

show abstract

“…T. brucei and L. mexicana have a single copy sequence, whereas T. cruzi has five copies, two of them being pseudogenes. The crystal structure of the TcG6PDH revealing the molecular basis of substrate and cofactor recognition was recently obtained showing that differences in the cofactor binding site compared to the human homologue allowing future exploration towards the design of new competitive NADP+ inhibitors [250].…”

Section: Oxidative Branchmentioning

confidence: 99%

Potential Drug Targets in the Pentose Phosphate Pathway of Trypanosomatids

Loureiro

Faria

Smith

et al. 2019

CMC

View full text Add to dashboard Cite

The trypanosomatids, Trypanosoma brucei, Trypanosoma cruzi and Leishmania spp, are causative agents of important human diseases such African sleeping sickness, Chagas' disease and Leishmaniasis, respectively. The high impact of these diseases on human health and economy worldwide, the unsatisfactory available chemotherapeutic options and the absence of human effective vaccines, strongly justifies the search for new drugs. The pentose phosphate pathway has been proposed to be a viable strategy to defeat several infectious diseases, including those from trypanosomatids, as it includes an oxidative branch, important in the maintenance of cell redox homeostasis, and a non-oxidative branch in which ribose 5-phosphate and erythrose 4-phosphate, precursors of nucleic acids and aromatic amino acids, are produced. This review provides an overview of the available chemotherapeutic options against these diseases and discusses the potential of genetically validated enzymes from the pentose phosphate pathway of trypanosomatids to be explored as potential drug targets.

show abstract

The structure of a Trypanosoma cruzi glucose‐6‐phosphate dehydrogenase reveals differences from the mammalian enzyme

Cited by 23 publications

References 42 publications

Biochemical Characterization and Structural Modeling of Fused Glucose-6-Phosphate Dehydrogenase-Phosphogluconolactonase from Giardia lamblia

Biochemical Characterization and Structural Modeling of Fused Glucose-6-Phosphate Dehydrogenase-Phosphogluconolactonase from Giardia lamblia

Cofactor specificity of glucose-6-phosphate dehydrogenase isozymes inPseudomonas putidareveals a general principle underlying glycolytic strategies in bacteria

Potential Drug Targets in the Pentose Phosphate Pathway of Trypanosomatids

Contact Info

Product

Resources

About