Biochemical properties of a purified protein in cystic fluid of Taenia solium metacestodes

Abstract

By affinity chromatography using a monoclonal antibody as ligand, Kim et al. (1986) purified a protein fraction in cystic fluid of Taenia solium metacestodes (CF). In this study, the biochemical properties of the purified protein were characterized. Discontinuous-polyacrylamide gel electrophoresis (disc-PAGE) of the protein at 4.5~10% separating gel concentration showed its molecular weight (MW) to be 150 kilodalton(kDa) in non-denatured state, while denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed that it was composed of 3 different subunits with respective MW of 15, 10 and 7 kDa. Subunit of 7 kDa was shown to be linked to other subunits by disulfide bonds. Isoelectric point of the protein was pH 6.8. The protein was relatively heat-stable for immunologic analysis. These properties indicated that the protein, comprising about 70% of total content in CF, had similar biochemical characters with antigen B of Oriol et al.(1971) in hydatid cyst fluid (HF)

Immunolocalization of the 150 kDa protein in cyst fluid of Taenia solium metacestodes

“…1), and this procedure correlated with the results of a previous study (Cho et al, 1988). The native molecular weight of the protein was estimated to be about 150 kDa by a Superose 6 HR 10/30 gel filtration, and the purified protein was soluble even at high concentration of ammonium sulfate solution (up to 80%).…”

Proteomic analysis of a 120 kDa protein complex in cyst fluid of Taenia solium metacestode and preliminary evaluation of its value for the serodiagnosis of neurocysticercosis

“…These purified GST fusion proteins were directly used in immunoblot assay because we found that the GST domain did not affect the antibody reactivity of the recombinant proteins (data not shown). A panel of low molecular weight proteins in TsM have previously been identified and/or purified, using affinity columns with either monoclonal antibodies Cho et al 1988) or lectin Lens calinaris (Tsang et al 1989 ;Restrepo et al 2000 ;Haslam et al 2003), or by preparative isoelectric focusing Ko and Ng, 1998). 7.…”

“…We previously described a highly antigenic and specific 150 kDa protein complex, which was composed of 3 subunits of 7, 10 and 15 kDa, in the TsM CF Cho et al 1988 ;Yang et al 1998 ;Chung et al 1999); however, it remains to be determined how many more macromolecular proteins exist in TsM other than the 150 kDa molecule, whether they consist of subunit proteins, and, if they are, the manner in which the subunits are linked, as well as which subunits are the most specific and useful in the context of serodiagnosis. We previously described a highly antigenic and specific 150 kDa protein complex, which was composed of 3 subunits of 7, 10 and 15 kDa, in the TsM CF Cho et al 1988 ;Yang et al 1998 ;Chung et al 1999); however, it remains to be determined how many more macromolecular proteins exist in TsM other than the 150 kDa molecule, whether they consist of subunit proteins, and, if they are, the manner in which the subunits are linked, as well as which subunits are the most specific and useful in the context of serodiagnosis.…”

Immunoblot findings of calcareous corpuscles binding proteins in cyst fluid of Taenia solium metacestodes

“…1B). Antigenic protein of 10 kDa is known to be a subunit of a 150 kDa protein in the cyst fluid of T. solium metacestodes and is a highly specific and sensitive antigen for the diagnosis of cysticercosis (Cho et al, 1988(Cho et al, , 1992Yang et al, 1998). This molecule has been cloned and its antigenic sites analyzed (Chung et al, 1999(Chung et al, , 2002.…”