Gene sequencing, modelling and immunolocalization of the protein disulfide isomerase from Plasmodium chabaudi.
Malaria remains one of the major human parasitic diseases, particularly in subtropical regions. Most of the fatal cases are caused by Plasmodium falciparum. The rodent parasite Plasmodium chabaudi has been the model of choice in research due to its similarities to human malaria, including developmental cycle, preferential invasion of mature erythrocytes, synchrony of asexual development, antigenic variation, gene sinteny as well as similar resistance mechanisms. Protein disulfide isomerase (PDI) is an essential catalyst of the endoplasmic reticulum in different biological systems with folding and chaperone activities. Most of the proteins exported by parasites have to pass through the endoplasmic reticulum before reaching their final destination and their correct folding is critical for parasite survival. PDI constitutes a potential target for the development of alternative therapy strategies based on the inhibition of folding and chaperoning of exported proteins. We here describe the sequencing of the gene coding for the PDI from P. chabaudi and analyse the relationship to its counterpart enzymes, particularly with the PDI from other Plasmodium species. The model constructed, based on the recent model deduced from the crystallographic structure 2B5E, was compared with the previous theoretical model for the whole PDI molecule constructed by threading. A recombinant PDI from P. chabaudi was also produced and used as an antigen for monoclonal antibody production for application in PDI immunolocalization.
Novo
Carlos
C
Unidade de Tecnologias de Proteínas e Anticorpos Monoclonais (UTPAM), Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Edifício F, Estrada do Paço do Lumiar, 1649-038 Lisboa, Portugal. cnovo@ihmt.unl.pt
Martins
Tiago M
TM
Prata
Sofia
S
Lopes
Angela
A
Armada
Ana
A
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D045744:Cell Line, Tumor; D018360:Crystallography, X-Ray; D005455:Fluorescent Antibody Technique; D016678:Genome / Q000235:genetics; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010802:Phylogeny; D016788:Plasmodium chabaudi / Q000201:enzymology*; D011487:Protein Conformation; D019704:Protein Disulfide-Isomerases / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D017422:Sequence Analysis, DNA
D000911:Antibodies, Monoclonal; D019704:Protein Disulfide-Isomerases