Branquinha M H, Bergter E B, de Meirelles M N, Vermelho A B
Departamento de Microbiologia Geral, Universidade Federal do Rio de Janeiro, Brasil.
Parasitol Res. 1994;80(4):336-41. doi: 10.1007/BF02351876.
A comparative study of glycolipid and protein composition in Trypanosoma cruzi and non-pathogenic trypanosomatids was carried out using Triton X-114 extraction. Protein profiles were analysed by polyacrylamide gel electrophoresis in sodium dodecyl sulphate (SDS-PAGE), and glycolipids were detected using high-performance thin-layer chromatography (HPTLC). Hydrophilic protein profiles were similar in non-pathogenic protozoa. Endotrypanum schaudinni, Crithidia luciliae and T. mega showed five characteristic protein bands ranging between 30 and 66 kDa. In the hydrophobic phase, a band of 50 kDa was present only in T. mega. Strain-specific protein distribution was detected in T. cruzi clone Dm28c and T. cruzi G and Y strains; clone Dm28c had five typical hydrophilic proteins at between 24 and 45 kDa, the G strain had two bands at 45 kDa in the hydrophilic phase and the Y strain had a major protein band at 24 kDa in both phases. T. dionisii and T. cruzi clone Dm28c showed a characteristic distribution of three hydrophilic proteins of approx. 45 kDa. Qualitative analysis of glycolipid composition showed that the T. cruzi strains and Dm28c clone and T. dionisii had four orcinol-positive spots, whereas in the other non-pathogenic trypanosomatids only three glycolipids were detected.
利用Triton X - 114萃取法,对克氏锥虫和非致病性锥虫的糖脂和蛋白质组成进行了比较研究。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS - PAGE)分析蛋白质谱,并用高效薄层色谱法(HPTLC)检测糖脂。非致病性原生动物的亲水性蛋白质谱相似。肖氏内锥虫、鲁氏克氏锥虫和巨型锥虫显示出五条特征性蛋白带,分子量在30至66 kDa之间。在疏水相中,仅巨型锥虫存在一条50 kDa的条带。在克氏锥虫克隆Dm28c以及克氏锥虫G株和Y株中检测到菌株特异性的蛋白质分布;克隆Dm28c在24至45 kDa之间有五条典型的亲水性蛋白,G株在亲水性相中45 kDa处有两条条带,Y株在两个相中均有一条主要的24 kDa蛋白条带。狄氏锥虫和克氏锥虫克隆Dm28c显示出三种约45 kDa亲水性蛋白的特征性分布。糖脂组成的定性分析表明,克氏锥虫各菌株、Dm28c克隆以及狄氏锥虫有四个苔黑酚阳性斑点,而在其他非致病性锥虫中仅检测到三种糖脂。