Eukaryotic translation initiation factor 5A and its posttranslational modifications play an important role in proliferation and potentially in differentiation of the human enteric protozoan parasite Entamoeba histolytica
Fig 8
SUnSET analysis of active protein synthesis.
(A) SUnSET analysis of wild type strain. Trophozoites of E. histolytica clonal strain HM-1:IMSS cl 6 were incubated in normal growth medium with 100 μg/ml cycloheximide, 10 μg/ml puromycin, or both. Cell lysates were subjected to SDS-PAGE and immunoblot using antibody specific for puromycin. Note that trophozoites incorporated puromycin into proteins (P). The incorporation was blocked by cycloheximide treatment (C + P). These data validated the SUnSET analysis for assessing protein synthesis in E. histolytica. (B) SUnSET analysis of EheIF5A2gs strain. Trophozoites from EheIF5A2gs and control (pSAP2G) strains were subjected to SUnSET analysis as in (A). Equal protein loads were confirmed with CS1 antibody (lower panels). Representative data for at least 3 separate trials are shown.