Fructose-bisphosphate aldolase and enolase from Echinococcus granulosus: Genes, expression patterns and protein interactions of two potential moonlighting proteins

• Published in: Gene Vol. 506; no. 1; pp. 76 - 84
• Main Authors: Lorenzatto, Karina Rodrigues, Monteiro, Karina Mariante, Paredes, Rodolfo, Paludo, Gabriela Prado, da Fonsêca, Marbella Maria, Galanti, Norbel, Zaha, Arnaldo, Ferreira, Henrique Bunselmeyer
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 10-09-2012
• Subjects: actin; Amino Acid Sequence; Animals; antibodies; Base Sequence; binding sites; Cestode; DNA, Helminth - genetics; Echinococcosis - parasitology; Echinococcus granulosus; Echinococcus granulosus - enzymology; Echinococcus granulosus - genetics; Echinococcus granulosus - pathogenicity; Echinococcus multilocularis; Echinococcus multilocularis - enzymology; Echinococcus multilocularis - genetics; fructose-bisphosphate aldolase; Fructose-Bisphosphate Aldolase - chemistry; Fructose-Bisphosphate Aldolase - genetics; Fructose-Bisphosphate Aldolase - metabolism; Gene Expression Profiling; genes; Genes, Helminth; glycolysis; Glycolytic enzymes; Helminth Proteins - chemistry; Helminth Proteins - genetics; Helminth Proteins - metabolism; Host-Parasite Interactions; Host–parasite interplay; Isoenzymes - chemistry; Isoenzymes - genetics; Isoenzymes - metabolism; larvae; Models, Molecular; Molecular Sequence Data; Moonlighting functions; parasites; Phosphopyruvate Hydratase - chemistry; Phosphopyruvate Hydratase - genetics; Phosphopyruvate Hydratase - metabolism; Protein Interaction Mapping; protoscoleces; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Sequence Homology, Amino Acid; Species Specificity; ORF; Host–parasite interplay; FBA; TEV; EST; IgG; Cestode; DAPI; Fru 1,6-P2; GST; ES; rEgEno1; LC-MS/MS; rEgFBA1; SDS-PAGE; Sulfo-SBED; Moonlighting functions; EgEno; IPTG; cDNA; Glycolytic enzymes; EgFBA; ELISA
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/j.gene.2012.06.046

Glycolytic enzymes, such as fructose-bisphosphate aldolase (FBA) and enolase, have been described as complex multifunctional proteins that may perform non-glycolytic moonlighting functions, but little is known about such functions, especially in parasites. We have carried out in silico genomic searches in order to identify FBA and enolase coding sequences in Echinococcus granulosus, the causative agent of cystic hydatid disease. Four FBA genes and 3 enolase genes were found, and their sequences and exon–intron structures were characterized and compared to those of their orthologs in Echinococcus multilocularis, the causative agent of alveolar hydatid disease. To gather evidence of possible non-glycolytic functions, the expression profile of FBA and enolase isoforms detected in the E. granulosus pathogenic larval form (hydatid cyst) (EgFBA1 and EgEno1) was assessed. Using specific antibodies, EgFBA1 and EgEno1 were detected in protoscolex and germinal layer cells, as expected, but they were also found in the hydatid fluid, which contains parasite's excretory–secretory (ES) products. Besides, both proteins were found in protoscolex tegument and in vitro ES products, further suggesting possible non-glycolytic functions in the host–parasite interface. EgFBA1 modeled 3D structure predicted a F-actin binding site, and the ability of EgFBA1 to bind actin was confirmed experimentally, which was taken as an additional evidence of FBA multifunctionality in E. granulosus. Overall, our results represent the first experimental evidences of alternative functions performed by glycolytic enzymes in E. granulosus and provide relevant information for the understanding of their roles in host–parasite interplay.
► There are four FBA and three enolase genes in E. granulosus and E. multilocularis. ► In E. granulosus only EgFBA1 and EgEno1 were detected in the pathogenic larval stage. ► EgFBA1 and EgEno1 proteins were detected in host–parasite interface compartments. ► EgFBA1 has a functional moonlighting actin binding site.