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Parasit Vectors. 2015 Sep 18;8:479. doi: 10.1186/s13071-015-1098-5.

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

Author information

1
Department of Anatomy, School of Medicine, University of Tampere, Tampere, Finland. zolfaghari.emameh.reza.x@student.uta.fi.
2
BioMediTech, University of Tampere, Tampere, Finland. zolfaghari.emameh.reza.x@student.uta.fi.
3
Fimlab Laboratories Ltd and Tampere University Hospital, Tampere, Finland. zolfaghari.emameh.reza.x@student.uta.fi.
4
Department of Anatomy, School of Medicine, University of Tampere, Tampere, Finland. marianne.kuuslahti@staff.uta.fi.
5
Dipartimento di Chimica, Laboratorio di Chimica Bioinorganica, Universita' degli Studi di Firenze, Sesto Fiorentino, Firenze, Italy. daniela.vullo@unifi.it.
6
Neurofarba Department, Sezione di Scienze Farmaceutiche e Nutraceutiche, Universita' degli Studi di Firenze, Sesto Fiorentino, Firenze, Italy. daniela.vullo@unifi.it.
7
Department of Anatomy, School of Medicine, University of Tampere, Tampere, Finland. Barker.Harlan.R@student.uta.fi.
8
Dipartimento di Chimica, Laboratorio di Chimica Bioinorganica, Universita' degli Studi di Firenze, Sesto Fiorentino, Firenze, Italy. claudiu.supuran@unifi.it.
9
Neurofarba Department, Sezione di Scienze Farmaceutiche e Nutraceutiche, Universita' degli Studi di Firenze, Sesto Fiorentino, Firenze, Italy. claudiu.supuran@unifi.it.
10
Department of Anatomy, School of Medicine, University of Tampere, Tampere, Finland. seppo.parkkila@staff.uta.fi.
11
Fimlab Laboratories Ltd and Tampere University Hospital, Tampere, Finland. seppo.parkkila@staff.uta.fi.

Abstract

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 kcat of 6.0 × 10(5) s(-1) and kcat/KM 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.

PMID:
26385556
PMCID:
PMC4575479
DOI:
10.1186/s13071-015-1098-5
[Indexed for MEDLINE]
Free PMC Article

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