Heterologous expression of a Trypanosoma cruzi surface glycoprotein (gp82) in mammalian cells indicates the existence of different signal sequence requirements and processing.
Metacyclic trypomastigotes of Trypanosoma cruzi express a developmentally regulated 82 kDa surface glycoprotein (gp82) that has been implicated in the mammalian cell invasion. When the non-infective epimastigote stage of the parasite was transfected with a vector containing the gp82 gene, an 82 kDa surface glycoprotein, which was indistinguishable from the metacyclic stage protein, was expressed. In contrast, when the same gene was expressed in transfected mammalian cells, although a large amount of protein was produced, it was not imported into the endoplasmic reticulum and glycosylated. This blockage in targeting and processing could be partially compensated for by the addition of a virus haemagglutinin signal peptide to the amino terminus of gp82. Thus, the requirements for membrane protein processing are distinct in mammals and T. cruzi, and an intrinsic feature of the gp82 prevents subsequent sorting to the mammalian cell surface. These results could be useful in the development of new DNA vaccines against T. cruzi employing parasite genes encoding immunodominant surface glycoproteins.
Ramirez
M I
MI
Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo, Escola Paulista de Medicina, Brazil.
Boscardin
S B
SB
Han
S W
SW
Paranhos-Baccala
G
G
Yoshida
N
N
Kelly
J M
JM
Mortara
R A
RA
Da Silveira
J F
JF
D000595:Amino Acid Sequence; D000818:Animals; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D004285:Dogs; D006031:Glycosylation; D008322:Mammals; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals; D014162:Transfection; D014349:Trypanosoma cruzi / Q000235:genetics*; D014709:Vero Cells
D008562:Membrane Glycoproteins; D021382:Protein Sorting Signals