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Identification, Molecular and Functional Characterization of Calmodulin Gene of Phytomonas serpens 15T that Shares High Similarity with its Pathogenic Counterparts Trypanosoma cruzi

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Abstract

In trypanosomatids, Ca2+-binding proteins can affect parasite growth, differentiation and invasion. Due to their importance for parasite maintenance, they become an attractive target for drug discovery and design. Phytomonas serpens 15T is a non-human pathogenic trypanosomatid that expresses important protein homologs of human pathogenic trypanosomatids. In this study, the coding sequence of calmodulin, a Ca2+-binding protein, of P. serpens 15T was cloned and characterized. The encoded polypeptide (CaMP) displayed high amino acid identity to homolog protein of Trypanosoma cruzi and four helix-loop-helix motifs were found. CaMP sequence analysis showed 20 amino acid substitutions compared to its mammalian counterparts. This gene is located on a chromosomal band with estimated size of 1,300 kb and two transcripts were detected by Northern blot analysis. A polyclonal antiserum raised against the recombinant protein recognized a polypeptide with an estimated size of 17 kDa in log-phase promastigote extracts. The recombinant CaMP retains its Ca2+-binding capacity.

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Abbreviations

CaMP:

Calmodulin from Phytomonas serpens 15T

dNTP:

2′ Deoxynucleoside-5′ triphosphate

EDTA:

Ethylene-diamine tetraacetic acid

EF:

Hand helix-loop-helix structural domain

EST:

Expressed sequence tag

GYPMI:

Glucose, yeast extract, peptone, meat infusion culture medium

IPTG:

Isopropyl-β-D-thiogalactopyranoside

SDS:

Sodium dodecyl sulphate

TBE:

Tris-borate-EDTA buffer

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Acknowledgments

We thank Paulo Lorca for help in camP nucleotide sequencing. We are also grateful to Dr. A. Leyva for reading this manuscript. This work was supported by grants from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Fundação Araucária and Pro-Reitoria de Pesquisa e Pós Graduação (PROPPG) of Universidade Estadual de Londrina (UEL). This study was part of the M.Sc. thesis work of T.A.C.B. Souza.

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Authors and Affiliations

  1. Departamento de Microbiologia, Centro de Ciências Biológicas, Universidade Estadual de Londrina, Rodovia Celso Garcia Cid, PR 445, km 380, Londrina, Paraná, CEP 86051-980, Brazil

    Tatiana de Arruda Campos Brasil de Souza, Viviane Krominski Graça-de Souza, César Armando Contreras Lancheros, Lucy Megumi Yamauchi & Sueli Fumie Yamada-Ogatta

  2. Instituto Carlos Chagas, Fundação Instituto Oswaldo Cruz, Curitiba, Paraná, Brazil

    Viviane Monteiro-Góes, Marco Aurélio Krieger & Samuel Goldenberg

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  1. Tatiana de Arruda Campos Brasil de Souza
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  2. Viviane Krominski Graça-de Souza
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  8. Sueli Fumie Yamada-Ogatta
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Correspondence to Sueli Fumie Yamada-Ogatta.

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de Arruda Campos Brasil de Souza, T., Graça-de Souza, V.K., Lancheros, C.A.C. et al. Identification, Molecular and Functional Characterization of Calmodulin Gene of Phytomonas serpens 15T that Shares High Similarity with its Pathogenic Counterparts Trypanosoma cruzi . Protein J 30, 212–219 (2011). https://doi.org/10.1007/s10930-011-9322-4

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  • DOI: https://doi.org/10.1007/s10930-011-9322-4

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