Selectivity of 3-bromo-isoxazoline inhibitors between human and Plasmodium falciparum glyceraldehyde-3-phosphate dehydrogenases

Bruno, Stefano; Margiotta, Marilena; Pinto, Andrea; Cullia, Gregorio; Conti, Paola; Micheli, C. De; Mozzarelli, Andrea

doi:10.1016/j.bmc.2016.04.033

Cited by 18 publications

(23 citation statements)

References 31 publications

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“…E2 enzymes are well-conserved in eukaryotes and involved in Ub/UBL-modification pathways, and play central roles in processes like regulating protein degradation, function, and localization, thereby controlling the biology of the eukaryotic cell [40]. GAPDH is a key glycolytic enzyme in the process of metabolism of coccidian, as several pathogenic protozoa entirely depend on glycolysis as the source of ATP in the host [41, 42]. Transhydrogenase catalyses transhydrogenation between analogues of NAD(H) and NADP(H).…”

Section: Discussionmentioning

confidence: 99%

“…GAPDH was proved to confer protection against Haemonchus contortus and Schistosoma mansoni [51, 52]. Protozoal GAPDHs were suggested as a potential antiparasitic targets in Plasmodium falciparum [41, 42], Leishmania mexicana [53], Trypanosoma brucei and Trypanosoma cruzi [54, 55]. Chen et al [56] reported that immunization with recombinant UCE induced protection against Taenia pisiformis .…”

Section: Discussionmentioning

confidence: 99%

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Identification of common immunodominant antigens of Eimeria tenella, Eimeria acervulina and Eimeria maxima by immunoproteomic analysis

et al. 2017

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Clinical chicken coccidiosis is mostly caused by simultaneous infection of several Eimeria species, and host immunity against Eimeria is species-specific. It is urgent to identify common immunodominant antigen of Eimeria for developing multivalent anticoccidial vaccines. In this study, sporozoite proteins of Eimeria tenella, Eimeria acervulina and Eimeria maxima were analyzed by two-dimensional electrophoresis (2DE). Western bot analysis was performed on the yielded 2DE gel using antisera of E. tenella E. acervulina and E. maxima respectively. Next, the detected immunodominant spots were identified by comparing the data from MALDI-TOF-MS/MS with available databases. Finally, Eimeria common antigens were identified by comparing amino acid sequence between the three Eimeria species. The results showed that analysis by 2DE of sporozoite proteins detected 629, 626 and 632 protein spots from E. tenella, E. acervulina and E. maxima respectively. Western bot analysis revealed 50 (E. tenella), 64 (E. acervulina) and 57 (E. maxima) immunodominant spots from the sporozoite 2DE gels of the three Eimeria species. The immunodominant spots were identified as 33, 27 and 25 immunodominant antigens of E. tenella, E. acervulina and E. maxima respectively. Fifty-four immunodominant proteins were identified as 18 ortholog proteins among the three Eimeria species. Finally, 5 of the 18 ortholog proteins were identified as common immunodominant antigens including elongation factor 2 (EF-2), 14-3-3 protein, ubiquitin-conjugating enzyme domain-containing protein (UCE) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). In conclusion, our results not only provide Eimeria sporozoite immunodominant antigen map and additional immunodominant antigens, but also common immunodominant antigens for developing multivalent anticoccidial vaccines.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Identification of common immunodominant antigens of Eimeria tenella, Eimeria acervulina and Eimeria maxima by immunoproteomic analysis

et al. 2017

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“…GAPDH is a key glycolytic enzyme in the process of metabolism of coccidian, as several pathogenic protozoa entirely depend on glycolysis as the source of ATP in the host. Thus, protozoal GAPDHs are considered potential targets for anti-protozoan drugs (Bruno et al, 2016, 2017). …”

Section: Introductionmentioning

confidence: 99%

Protective Efficacy of Coccidial Common Antigen Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) against Challenge with Three Eimeria Species

Tian

Huang

et al. 2017

Front. Microbiol.

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Coccidiosis is an intestinal disorder of poultry and often caused by simultaneous infections of several Eimeria species. GAPDH is one of the immunogenic common antigens among Eimeria tenella, E. acervulina, and E. maxima identified in our previous study. The present study was performed to further evaluate its immunogenicity and protective efficacy. The genes of GAPDH cloned from E. acervulina and E. maxima were named as EaGAPDH and EmGAPDH, respectively. The immunogenicity of recombinant proteins of EaGAPDH and EmGAPDH were analyzed by Western blot. The transcription and expression of pVAX-EaGAPDH and pVAX-EmGAPDH in the injected muscles were detected by reverse transcription PCR (RT-PCR) and Western blot, respectively. GAPDH-induced changes of T lymphocytes subpopulation, cytokines production, and antibody were determined using flow cytometry, quantitative real-time PCR (qPCR), and ELISA, respectively. Finally, the protective efficacies of pVAX-EaGAPDH and pVAX-EmGAPDH were evaluated by vaccination and challenge experiments. The results revealed that the recombinant GAPDH proteins reacted with the corresponding chicken antisera. The EaGAPDH genes were successfully transcribed and expressed in the injected muscles. Vaccination with pVAX-EaGAPDH and pVAX-EmGAPDH significantly increased the proportion of CD4+ and CD8+ T lymphocytes, the cytokines productions of IFN-γ, IL-2, IL-4 et al., and IgG antibody levels compared to controls. The vaccination increased the weight gains, decreased the oocyst outputs, alleviate the enteric lesions compared to controls, and induced moderate anti-coccidial index (ACI). In conclusion, the coccidial common antigen of GAPDH induced significant humoral and cellular immune response and effective protection against E. tenella, E. acervulina, E. maxima, and mixed infection of the three Eimeria species.

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“…The low pK a reflects the involvement of the catalytic cysteine in a cysteine–histidine catalytic dyad, which provides a pK a ‐lowering microenvironment that stabilizes the thiolate species . Consistently with its distinctive pK a , the catalytic cysteine of GAPDHs was shown to react with several chemically diverse electrophiles, such as iodoacetic acid, acrylonitrile, N ‐acetyl‐ p ‐benzoquinone imine, vinyl sulfones, 9,10‐phenanthrenequinone, and 3‐bromo‐isoxazoline …”

Section: Introductionmentioning

confidence: 93%

“…Particularly, GAPDHs from protozoan parasites have been validated as suitable molecular targets for antiparasitic drugs, as several pathogenic protozoa entirely depend on glycolysis as the source of ATP in the host stage. For instance, the absence of the pyruvate dehydrogenase complex in the mitochondria of Plasmodium falciparum ( Pf ) restricts its ATP production to glycolysis, an observation that prompted the structure‐based drug design of Pf GAPDH inhibitors . The unusual compartmentalization of glycolysis inside the specialized organelle glycosome in Trypanosoma and Leishmania suggested the development of GAPDH inhibitors for the treatment of Trypanosomatidae diseases .…”

Section: Introductionmentioning

confidence: 99%

Molecular basis for covalent inhibition of glyceraldehyde‐3‐phosphate dehydrogenase by a 2‐phenoxy‐1,4‐naphthoquinone small molecule

et al. 2017

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Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has recently gained attention as an antiprotozoan and anticancer drug target. We have previously identified 2-phenoxy-1,4-naphthoquinone as an inhibitor of both Trypanosoma brucei and human GAPDH. Herein, through multiple chemical, biochemical, and biological studies, and through the design of analogs, we confirmed the formation of a covalent adduct, we clarified the inhibition mechanism, and we demonstrated antitrypanosomal, antiplasmodial, and cytotoxic activities in cell cultures. The overall results lent support to the hypothesis that 2-phenoxy-1,4-naphthoquinone binds the GAPDH catalytic cysteine covalently through a phenolate displacement mechanism. By investigating the reactivity of 2-phenoxy-1,4-naphthoquinone and its analogs with four GAPDH homologs, we showed that the covalent inhibition is not preceded by the formation of a strong non-covalent complex. However, an up to fivefold difference in inactivation rates among homologs hinted at structural or electrostatic differences of their active sites that could be exploited to further design kinetically selective inhibitors. Moreover, we preliminarily showed that 2-phenoxy-1,4-naphthoquinone displays selectivity for GAPDHs over two other cysteine-dependent enzymes, supporting its suitability as a warhead starting fragment for the design of novel inhibitors.

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Selectivity of 3-bromo-isoxazoline inhibitors between human and Plasmodium falciparum glyceraldehyde-3-phosphate dehydrogenases

Cited by 18 publications

References 31 publications

Identification of common immunodominant antigens of Eimeria tenella, Eimeria acervulina and Eimeria maxima by immunoproteomic analysis

Identification of common immunodominant antigens of Eimeria tenella, Eimeria acervulina and Eimeria maxima by immunoproteomic analysis

Protective Efficacy of Coccidial Common Antigen Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) against Challenge with Three Eimeria Species

Molecular basis for covalent inhibition of glyceraldehyde‐3‐phosphate dehydrogenase by a 2‐phenoxy‐1,4‐naphthoquinone small molecule

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