Molecular cloning and characterization of a thioredoxin gene from Echinococcus granulosus.
The insert of a clone from a lambdagt11 Echinococcus granulosus (Platyhelminth, Cestoda) protoscolex cDNA library, showed an open reading frame whose deduced protein sequence presents a high homology with all described thioredoxins (TRX). The TRX active site (Trp-Cys-Gly-Pro-Cys) is completely conserved. With a monospecific antibody, selected from a total anti-protoscolex sera by the isolated clone, a 12 kDa polypeptide was immunoprecipitated from a protoscolex total protein extract. Furthermore, an antiserum raised against a recombinant EgTRX also recognizes a 12 kDa band in these extracts. The recombinant protein presents TRX activity, using the insulin reduction assay. Finally, a TRX activity was characterized in protoscolex extracts. In all organisms where TRXs were studied, they participate in a cascade of redox exchanges, contributing to the maintaining of cell homeostasis. Considering that the parasitic flatworm E. granulosus is probably submitted to an important oxidative stress due to host defences, EgTRX protein could be involved in the survival strategies of this parasite.
Chalar
C
C
Facultad de Ciencias, Universidad de la República, Iguá 4225, Montevideo, 11500, Uruguay. cora@fcien.edu.uy
Martínez
C
C
Agorio
A
A
Salinas
G
G
Soto
J
J
Ehrlich
R
R
D000595:Amino Acid Sequence; D000818:Animals; D015139:Blotting, Southern; D015153:Blotting, Western; D003001:Cloning, Molecular; D004446:Echinococcus / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D015870:Gene Expression; D015801:Helminth Proteins / Q000737:chemistry / Q000145:classification / Q000235:genetics / Q000378:metabolism; D007106:Immune Sera; D007328:Insulin / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames / Q000235:genetics; D010084:Oxidation-Reduction; D017433:Protein Structure, Secondary; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D011994:Recombinant Proteins / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013879:Thioredoxins / Q000737:chemistry / Q000145:classification / Q000235:genetics* / Q000378:metabolism*
D015801:Helminth Proteins; D007106:Immune Sera; D007328:Insulin; D012333:RNA, Messenger; D011994:Recombinant Proteins; D013879:Thioredoxins