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Ascaris lumbricoides β carbonic anhydrase: a potential target enzyme for treatment of ascariasis

BACKGROUND: A parasitic roundworm, Ascaris lumbricoides, is the causative agent of ascariasis, with approximately 760 million cases around the world. Helminthic infections occur with a high prevalence mostly in tropical and developing xcountries. Therefore, design of affordable broad-spectrum anti-h...

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Autores principales: Zolfaghari Emameh, Reza, Kuuslahti, Marianne, Vullo, Daniela, Barker, Harlan R., Supuran, Claudiu T., Parkkila, Seppo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4575479/
https://www.ncbi.nlm.nih.gov/pubmed/26385556
http://dx.doi.org/10.1186/s13071-015-1098-5
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author Zolfaghari Emameh, Reza
Kuuslahti, Marianne
Vullo, Daniela
Barker, Harlan R.
Supuran, Claudiu T.
Parkkila, Seppo
author_facet Zolfaghari Emameh, Reza
Kuuslahti, Marianne
Vullo, Daniela
Barker, Harlan R.
Supuran, Claudiu T.
Parkkila, Seppo
author_sort Zolfaghari Emameh, Reza
collection PubMed
description BACKGROUND: A parasitic roundworm, Ascaris lumbricoides, is the causative agent of ascariasis, with approximately 760 million cases around the world. Helminthic infections occur with a high prevalence mostly in tropical and developing xcountries. Therefore, design of affordable broad-spectrum anti-helminthic agents against a variety of pathogens, including not only A. lumbricoides but also hookworms and whipworms, is desirable. Beta carbonic anhydrases (β-CAs) are considered promising targets of novel anthelminthics because these enzymes are present in various parasites, while completely absent in vertebrates. METHODS: In this study, we identified an A. lumbricoides β-CA (AIBCA) protein from protein sequence data using bioinformatics tools. We used computational biology resources and methods (including InterPro, CATH/Gene3D, KEGG, and METACYC) to analyze AlBCA and define potential roles of this enzyme in biological pathways. The AlBCA gene was cloned into pFastBac1, and recombinant AIBCA was produced in sf-9 insect cells. Kinetics of AlBCA were analyzed by a stopped-flow method. RESULTS: Multiple sequence alignment revealed that AIBCA contains the two sequence motifs, CXDXR and HXXC, typical for β-CAs. Recombinant AIBCA showed significant CA catalytic activity with k(cat) of 6.0 × 10(5) s(−1) and k(cat)/K(M) of 4.3 × 10(7) M(−1) s(−1). The classical CA inhibitor, acetazolamide, showed an inhibition constant of 84.1 nM. Computational modeling suggests that the molecular architecture of AIBCA is highly similar to several other known β-CA structures. Functional predictions suggest that AIBCA might play a role in bicarbonate-mediated metabolic pathways, such as gluconeogenesis and removal of metabolically produced cyanate. CONCLUSIONS: These results open new avenues to further investigate the precise functions of β-CAs in parasites and suggest that novel β-CA specific inhibitors should be developed and tested against helminthic diseases. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13071-015-1098-5) contains supplementary material, which is available to authorized users.
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spelling pubmed-45754792015-09-20 Ascaris lumbricoides β carbonic anhydrase: a potential target enzyme for treatment of ascariasis Zolfaghari Emameh, Reza Kuuslahti, Marianne Vullo, Daniela Barker, Harlan R. Supuran, Claudiu T. Parkkila, Seppo Parasit Vectors Research BACKGROUND: A parasitic roundworm, Ascaris lumbricoides, is the causative agent of ascariasis, with approximately 760 million cases around the world. Helminthic infections occur with a high prevalence mostly in tropical and developing xcountries. Therefore, design of affordable broad-spectrum anti-helminthic agents against a variety of pathogens, including not only A. lumbricoides but also hookworms and whipworms, is desirable. Beta carbonic anhydrases (β-CAs) are considered promising targets of novel anthelminthics because these enzymes are present in various parasites, while completely absent in vertebrates. METHODS: In this study, we identified an A. lumbricoides β-CA (AIBCA) protein from protein sequence data using bioinformatics tools. We used computational biology resources and methods (including InterPro, CATH/Gene3D, KEGG, and METACYC) to analyze AlBCA and define potential roles of this enzyme in biological pathways. The AlBCA gene was cloned into pFastBac1, and recombinant AIBCA was produced in sf-9 insect cells. Kinetics of AlBCA were analyzed by a stopped-flow method. RESULTS: Multiple sequence alignment revealed that AIBCA contains the two sequence motifs, CXDXR and HXXC, typical for β-CAs. Recombinant AIBCA showed significant CA catalytic activity with k(cat) of 6.0 × 10(5) s(−1) and k(cat)/K(M) of 4.3 × 10(7) M(−1) s(−1). The classical CA inhibitor, acetazolamide, showed an inhibition constant of 84.1 nM. Computational modeling suggests that the molecular architecture of AIBCA is highly similar to several other known β-CA structures. Functional predictions suggest that AIBCA might play a role in bicarbonate-mediated metabolic pathways, such as gluconeogenesis and removal of metabolically produced cyanate. CONCLUSIONS: These results open new avenues to further investigate the precise functions of β-CAs in parasites and suggest that novel β-CA specific inhibitors should be developed and tested against helminthic diseases. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13071-015-1098-5) contains supplementary material, which is available to authorized users. BioMed Central 2015-09-18 /pmc/articles/PMC4575479/ /pubmed/26385556 http://dx.doi.org/10.1186/s13071-015-1098-5 Text en © Zolfaghari Emameh et al. 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Zolfaghari Emameh, Reza
Kuuslahti, Marianne
Vullo, Daniela
Barker, Harlan R.
Supuran, Claudiu T.
Parkkila, Seppo
Ascaris lumbricoides β carbonic anhydrase: a potential target enzyme for treatment of ascariasis
title Ascaris lumbricoides β carbonic anhydrase: a potential target enzyme for treatment of ascariasis
title_full Ascaris lumbricoides β carbonic anhydrase: a potential target enzyme for treatment of ascariasis
title_fullStr Ascaris lumbricoides β carbonic anhydrase: a potential target enzyme for treatment of ascariasis
title_full_unstemmed Ascaris lumbricoides β carbonic anhydrase: a potential target enzyme for treatment of ascariasis
title_short Ascaris lumbricoides β carbonic anhydrase: a potential target enzyme for treatment of ascariasis
title_sort ascaris lumbricoides β carbonic anhydrase: a potential target enzyme for treatment of ascariasis
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4575479/
https://www.ncbi.nlm.nih.gov/pubmed/26385556
http://dx.doi.org/10.1186/s13071-015-1098-5
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