Echinostoma caproni: identification of enolase in excretory/secretory products, molecular cloning, and functional expression.
In order to investigate molecules that could be involved in host-trematode relationships, we have analysed the excretory/secretory products (ESP) of Echinostoma caproni following a proteomic approach. Actin, Gluthathione S-transferase (GST) and enolase have been identified in the ESP. Enolase, observed to be one of the most abundant proteins, was further characterized. The molecular cloning and in vitro expression in Escherichia coli of E. caproni enolase allowed us to determine that the protein contains 431 amino acids and a theoretical MW of 46272 Da. E. caproni enolase shows high homology to other trematode enolases. The recombinant protein binds specifically to human plasminogen in vitro, as observed for the native protein, confirming its properties as a host-interacting molecule.
Marcilla
Antonio
A
Departamento de Biología Celular y Parasitologia, Facultat de Farmàcia, Universitat de Valencia, Av. V.A. Estellés, s/n, 46100 Burjassot, Valencia, Spain. Antonio.marcilla@uv.es
Pérez-García
Ana
A
Espert
Ana
A
Bernal
Dolores
D
Muñoz-Antolí
Carla
C
Esteban
José Guillermo
JG
Toledo
Rafael
R
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D003001:Cloning, Molecular; D006224:Cricetinae; D004449:Echinostoma / Q000201:enzymology* / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D008297:Male; D008647:Mesocricetus; D008969:Molecular Sequence Data; D010751:Phosphopyruvate Hydratase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D010958:Plasminogen / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D011994:Recombinant Proteins; D010958:Plasminogen; D010751:Phosphopyruvate Hydratase