Abstract
The spectrum of proteins was studied in plerocercoids of parasitic cestodes Triaenophorus nodulosus collected from the liver of different species of fish: perch Perca fluviatilis L ., ruffe Gymnocephalus cernuus L. and burbot Lota lota L. A comparison of protein extracts separated by 2D electrophoresis revealed 18 proteins whose concentrations differed among larvae of different fish species. Of these, 4 proteins were identified by using mass-spectrometric methods (MALDI-TOF/TOF, LC–MS): a carbohydrate metabolism enzyme triosephosphate isomerase, the proteins of microtubules tubulin α and β, and the G-protein signaling molecule. The elevated content of components of the G-protein pathway of the cytoskeleton and cell morphology regulation (tubulin and G-protein) in plerocercoids from perch in comparison with plerocercoids from burbot indicate a more dynamic cytoskeleton, increased cell proliferation and differentiation processes, and possibly more mature larvae in the former species. A higher level of expression of the key enzyme of glycolysis and signal transmission in plerocercoids from perch compared to plerocercoids from burbot indicates a greater metabolic activity in larvae from perch. The obtained results indicate that the expression of some proteins in T. nodulosus larvae is influenced by the habitat.
Highlights
The spectrum of proteins was studied in plerocercoids of parasitic cestodes Triaenophorus nodulosus collected from the liver of different species of fish: perch Perca fluviatilis L., ruffe Gymnocephalus cernuus L. and burbot Lota lota L
4 proteins were identified by using mass-spectrometric methods (MALDI-TOF/TOF, LC–MS): a carbohydrate metabolism enzyme triosephosphate isomerase, the proteins of microtubules tubulin α and β, and the G-protein signaling molecule
The obtained results indicate that the expression of some proteins in T. nodulosus larvae is influenced by the habitat
Summary
Плероцеркоиды гельминтов были собраны из рыб, выловленных из водоемов Республики Карелия: в сентябре 2015 г. в Ладожском озере (окунь Perca fluviatilis, ерш Gymnocephalus cernuus); в феврале-марте 2016 г. в Онежском озере (налим Lota lota). В день анализа очищенный белок растворяли в буфере, содержащем 8 M мочевину, 2% Triton X-100, 0,05 M дитиотриетол, 0,5% амфолиты с диапазоном рН 3,5–10 (BioRad) с добавлением коктейля ингибиторов протеаз и фенилметансульфонилфторида (до конечных концентраций в растворе 1 %) [O'Farrell, 1975]. Изоэлектрофокусирование проводили при помощи модифицированной системы BioRad Protean II Xi cell (США) в течение 18 ч, постепенно повышая силу тока от 100 до 900 мА. Затем гели извлекали из капилляров и инкубировали в растворе Лэммли (9% SDS, 6% β-меркаптоэтанол (объем/объем), 10% глицерин (объем/объем) с добавлением 2–5 мг бромфенолового синего в 0,2 M ТрисHCl, pH 6,7). Полученные после изофокусирования, накладывали на 10% ПААГ-гели и проводили денатурирующий электрофорез по Лэммли [Laemmli, 1970] в ячейке системы BioRad Protean II Xi cell (США) с охлаждением. После проведения электрофореза гели были окрашены в течение 30 мин в растворе 0,2 % Кумасси G-250 в 25% изопропаноле и 10% уксусной кислоте. Окрашенные гели фотографировали при помощи системы гель-документации MiniLumi (Berthold Technologies)
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