Abstract
Among the microbodies found in eukaryotes are the glycosomes of Trypanosoma brucei, thought to be closely related to peroxisomes. Two types of targeting signals for glycosomes have been identified thus far: type 1 at the C-terminus and type 2 at the N-terminus. In this report, we use an epitope-tagging system to characterize the targeting signal found on the minor glycosomal isozyme of phosphoglycerate kinase, 56PGK. No type 1 or 2 signal was found; rather, the topogenic information was found to be internal. Chimeric molecules formed with the cytoplasmic phosphoglycerate kinase isozyme indicate that a region between amino acids 24 and 91 of 56PGK is essential for glycosomal targeting. No homology was found between this region and peroxisomal proteins containing internal targeting signals.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Animals
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Cytoplasm / enzymology
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Epitopes / chemistry
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Epitopes / genetics
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Fluorescent Antibody Technique
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Immunoblotting
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Isoenzymes / analysis*
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Isoenzymes / chemistry
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Isoenzymes / genetics
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Microbodies / enzymology*
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Molecular Sequence Data
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Mutagenesis
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Phosphoglycerate Kinase / analysis*
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Phosphoglycerate Kinase / chemistry
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Phosphoglycerate Kinase / genetics
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Plasmids
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Proto-Oncogene Proteins c-myc / chemistry
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Proto-Oncogene Proteins c-myc / genetics
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics
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Transfection
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Trypanosoma brucei brucei / enzymology*
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Trypanosoma brucei brucei / genetics
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Trypanosoma brucei brucei / ultrastructure
Substances
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Epitopes
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Isoenzymes
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Proto-Oncogene Proteins c-myc
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Recombinant Fusion Proteins
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Phosphoglycerate Kinase