A rapid method for the isolation of intact glycosomes from Trypanosoma brucei by Percoll -gradient centrifugation in a vertical rotor.
A rapid method for the isolation of intact glycosomes from trypanosoma brucei is described. A post large-granule extract is subjected to density-gradient centrifugation on isotonic Percoll in a vertical rotor. This results in a good separation of the glycosomes, which equilibrated at 1.09 g/cm3, from other cellular components. Glycosomes isolated from such a gradient are 9-fold purified relative to the starting homogenate with a yield of 25%. Their glycolytic enzymes exhibit a high latency indicative of an intact glycosomal membrane.
D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D002499:Centrifugation, Density Gradient; D005956:Glucose-6-Phosphate Isomerase / Q000378:metabolism; D006019:Glycolysis; D006593:Hexokinase / Q000378:metabolism; D009940:Organoids / Q000201:enzymology*; D011205:Povidone; D012822:Silicon Dioxide; D014346:Trypanosoma brucei brucei / Q000201:enzymology / Q000648:ultrastructure*
C016039:Percoll; D012822:Silicon Dioxide; D006593:Hexokinase; D005956:Glucose-6-Phosphate Isomerase; D011205:Povidone