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.
Departamento de Imunologia, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brazil.
R T
RT
Pinho
H S
HS
Dutra
S
S
Giovanni-De-Simone
L C
LC
de Carvalho
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