Taenia solium: identification and preliminary characterization of a lipid binding protein with homology to the SEC14 catalytic domain |
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Authors: | Montero Estrella Gonzalez Luis Miguel Bonay Pedro Rosas Gabriela Hernandez Beatriz Sciutto Edda Parkhouse R Michael E Harrison Leslie J S Morales Miguel Angel Garate Teresa |
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Affiliation: | Instituto de Salud Carlos III, Centro Nacional de Microbiología, Ctra Majadahonda Pozuelo Km 2,2, 28220, Majadahonda Madrid, Spain. |
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Abstract: | The objective of this work is to identify proteins of the human and porcine parasite, Taenia solium, which may be exploited for control of the parasite. Through screening a cDNA library of T. solium metacestodes, we have identified a novel Sec-14-like Taenia lipid-binding protein that may play an important role in membrane trafficking. The Sec14-like sequence is a single copy gene, encoding a putative polypeptide of 320 amino acids and 36.1 kDa (sec14Tsol protein). Secondary amino acid structural analysis suggested that the sec14Tsol protein might contain two distinct structural domains, an amino-terminal alpha-helix rich domain and a mixed alpha-helix/beta-stand carboxy-terminal zone, showing homology with the conserved SEC14 domain found in a great number of proteins that bind lipids, as the regulators of membrane trafficking between Golgi membrane bilayers. Significantly, therefore, in a phosphoinositide-binding assay, sec14Tsol purified recombinant protein specifically interacted with important lipid regulators of membrane trafficking, with a preference for PI(3)P(2), PI(3,4)P(2), PI(4,5)P(2) and phosphatidic acid. Moreover, the sec14Tsol protein was localized in the Golgi apparatus of transfected cells and in the spiral canal region of T. solium metacestode tegument. As sec14Tsol protein may play an important role in membrane trafficking, its demonstrated localisation in the intact parasite tegument suggests its involvement in the function of the tegument and thus perhaps interaction with the host. |
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Keywords: | SEC14 domain Sec14-like gene Lipid binding Taenia solium BCA, bicinchoninic acid bp, base pair BSA, bovine serum albumin cDNA, copy deoxyribonucleic acid EDTA, ethylenediaminetetraacetic acid FITC, fluorescein isothiocyanate Gal-T, beta 1-4-galactosyl-transferase gDNA, genomic deoxyribonucleic acid Kb, kilobase kDA, kiloDalton LPA, lysophosphatidic acid LPC, lysophosphacholine NRK, normal rat kidney PA, phosphatidic acid PBS, phosphate buffered saline PBS/A-T, 1% BSA in PBS plus 0,1% Triton X-100 ORF open reading frame PC, phosphatidylcholine PCR, polymerase chain reaction PE, phosphatidylethanolamine PI, phosphatidylinositol PIP, phosphatidylinositol phosphate PS, phosphatidyl serine S1P sphingosine-1-phosphate SEC14, S. cerevisiae gene. |
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