A glass wool-based method for purifying Trypanosoma cruzi trypomastigotes and identification of an epimastigote-specific glass-adherent surface peptide.
Glass wool, hydrophilic cotton wool, non-electrically charged BIO-GEL P2 and common tissue paper columns were used to purify trypomastigotes from a mixed Trypanosoma cruzi population grown in axenic culture medium. With all these columns, highly purified (up to 98%) trypomastigote preparations were obtained. Trypomastigote yields from cotton wool, BIO-GEL P2 and common tissue paper columns were not as high as from glass wool columns, from which yields varied from 69 to 80%. Purification on glass wool did not affect trypomastigote infectivity or virulence. Dead trypomastigotes could not be purified on glass wool columns. A glass-adherent amphiphilic peptide of 45 kDa, present in the cell membrane, was isolated from epimastigote but not from trypomastigote preparations.
Pinho
R T
RT
Departamento de Imunologia, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brazil.
Dutra
H S
HS
Giovanni-De-Simone
S
S
de Carvalho
L C
LC
D000818:Animals; D014355:Chagas Disease / Q000469:parasitology*; D002848:Chromatography, DEAE-Cellulose; D004591:Electrophoresis, Polyacrylamide Gel; D005898:Glass; D007303:Insect Vectors / Q000469:parasitology; D008297:Male; D008565:Membrane Proteins / Q000032:analysis* / Q000378:metabolism; D051379:Mice; D015800:Protozoan Proteins / Q000032:analysis* / Q000378:metabolism; D014224:Triatoma / Q000469:parasitology; D014349:Trypanosoma cruzi / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
D008565:Membrane Proteins; D015800:Protozoan Proteins