Reversed phase HPLC of different membrane-bound variant surface glycoprotein preparations from Trypanosoma brucei brucei.

Abstract

The chromatographic behaviour of the membrane-attached variant surface glycoprotein (mfVSG) of bloodstream forms of Trypanosoma brucei brucei AnTat 1.1 A preparations were studied by reversed phase high performance liquid chromatography (RP-HPLC). Among the different preparation procedures used, only the trifluoroacetic acid extraction gave a mfVSG preparation which was eluted from the RP-HPLC column. As well as for the soluble variant surface glycoprotein (sVSG) it was found that different mfVSG forms were eluted at different organic solvent concentrations from the RP-HPLC column. In addition, in preliminary studies, we have attempted to characterize the factors responsible for the trypanosome surface coat assembly. Using chromatographic techniques (RP-HPLC and thin layer chromatography), the results suggest a close relationship between the content of lipid-containing species and the heterogeneity of mfVSG preparation.