[Determination of free thiols in the chimeric protein PfCP-2.9 of Plasmodium falciparum].
OBJECTIVE
To determine the free thiols in the chimeric protein PfCP-2.9 of Plasmodium falciparum expressed by Pichia pastoris.


METHODS
Two experiments of reverse phase HPLC and Ellman's reaction were applied to the PfCP-2.9 for the determination of its free thiols. For RP-HPLC analysis, three kinds of samples were tested: PfCP-2.9, dithiothreitol-reduced PfCP-2.9 and indoacetic acid-alkylated PfCP-2.9.


RESULTS
Both experiments showed that there were no any free thiols present in the PfCP-2.9.


CONCLUSIONS
The disulfide bonds between cysteine residues of PfCP-2.9 were formed completely.
Qian
Feng
F
Department of Etiological Biology, Second Military Medical University, Shanghai 200433.
Pan
Wei-qing
WQ
D000953:Antigens, Protozoan / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D008565:Membrane Proteins / Q000032:analysis*; D020066:Merozoite Surface Protein 1 / Q000032:analysis*; D015800:Protozoan Proteins / Q000032:analysis*; D011993:Recombinant Fusion Proteins / Q000032:analysis*; D013053:Spectrophotometry; D013438:Sulfhydryl Compounds / Q000032:analysis* / Q000737:chemistry
D000953:Antigens, Protozoan; D008565:Membrane Proteins; D020066:Merozoite Surface Protein 1; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; D013438:Sulfhydryl Compounds; C090077:apical membrane antigen I, Plasmodium