Molecular cloning and biochemical characterization of a VCP homolog in African trypanosomes.
作者： Janet L Roggy , James D Bangs
DOI: 10.1016/S0166-6851(98)00114-5
关键词:
摘要: Through reverse transcription-polymerase chain reaction using degenerate oligonucleotide primers, a VCP homolog was identified in African trypanosomes. Sequence analysis shows 72 and 64% deduced amino acid identity, respectively, with mouse yeast Cdc48p. Southern indicates tbVCP to have single locus two alleles. Antibodies generated against recombinant protein recognize 95 kDa whole cell lysates of both procyclic bloodstream There is an approximately four-fold greater expression TbVCP the stage trypanosome life cycle. Subcellular fractionation immunofluorescence anti-TbVCP antibodies indicate majority be cytoplasmically localized small subset associated membranes. Sucrose velocity sedimentation gel filtration size studies suggest that homohexameric particle as has been demonstrated other homologs. Also like homologs, contains NEM-inhibitable ATPase activity. This first characterization AAA (ATPases Associated variety cellular Activities) family member
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