Analysis of trypanosomal endocytic organelles using preparative free-flow electrophoresis.

摘要: In this paper we demonstrate the power of preparative free-flow electrophoresis (FFE) for study endocytosis by African trypanosomes. Endocytosis extracellular macromolecules these parasites occurs through a specialized region parasite called flagella pocket. The uptake fluid phase markers such as horseradish peroxidase (HRP) into various compartments endocytic pathway bloodstream forms Trypanosoma brucei was manipulated regulating external environment (e.g., altering temperature incubation). subcellular were then separated or isopycnic density gradient centrifugation and analyzed marker uptake. At low temperatures, HRP found predominantly in flagellar Increasing resulted time-dependent more positively charged endosomal fractions. However, little activity detected lysosomal compartments, suggesting that either had not yet entered lysosome degraded immediately upon entry. Through use FFE able to identify analyze possible alone. Although differences separation seen striking, minor changes within system subtle, depicted protease profiles. conclusion, show is powerful technique analysis pocket-derived membranes from other