Studies on the glycosome of Trypanosoma brucei
Glycosomes (microbodies) have been purified from bloodstream form Trypanosoma brucei by an improved procedure involving freezing and thawing live organisms in 15% glycerol prior to cell disruption. Highly purified organelles of bloodstream form T. brucei contain 11 major proteins of which 8 tentatively identified glycolytic enzymes make up about 90% of the total glycosomal protein. Treatment of these intact isolated organelles with the bisimidoester dimethylsuberimidate (DMSI) resulted in crosslinking of all glycosomal proteins into a large complex suggestive of juxtapositioning of the glycosomal proteins. The crosslinked complex was capable of catalyzing the multienzyme conversion of glucose to glycerol-3-phosphate but did not possess any special kinetic features different from those of the unaggregated enzymes represented by solubilized glycosomes. The multienzyme reaction had a lab phase associated with it and (/sup 14/C)-glucose label incorporation into sugar phosphate intermediates was effectively competed by unlabeled intermediates. Glycosomes were also purified from culture form T. brucei by several different procedures. Comparison of highly purified organelles from the two different life stages of the organism showed reduced specific activities and contents of the early glycolytic enzymes in organelles from the culture form with a decrease from 87% to 35% of the contribution of glycolytic enzymes to the total glycosomal protein.
- Research Organization:
- California Univ., San Francisco (USA)
- OSTI ID:
- 6937751
- Resource Relation:
- Other Information: Thesis (Ph. D.)
- Country of Publication:
- United States
- Language:
- English
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GLUCOSE
UPTAKE
ORGANOIDS
PURIFICATION
TRYPANOSOMES
ENZYMES
PROTEINS
BIOCHEMICAL REACTION KINETICS
CARBON 14 COMPOUNDS
CELL CULTURES
CELL MEMBRANES
GLYCOSYL HYDROLASES
TRACER TECHNIQUES
ALDEHYDES
CARBOHYDRATES
CELL CONSTITUENTS
HEXOSES
HYDROLASES
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
MEMBRANES
MONOSACCHARIDES
ORGANIC COMPOUNDS
REACTION KINETICS
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550701* - Microbiology- Tracer Techniques