Abstract
Ribosomal subunit protein 9 (rps9) is a nuclearly encoded protein that resides in the apicoplast organelle of Toxoplasma gondii. Two cis-acting regions within the rps9 transit domain (amino acids 38-49 and 79-86), when combined with the rps9 signal sequence, were necessary and sufficient for apicoplast targeting. To investigate proteins interacting with the rps9 leader sequence, parasites expressing rps9 leader constructs fused to a glutathione S-transferase (GST) reporter were prepared, and proteins associated with the leader constructs were purified from extracts by affinity chromatography. In addition to GST-containing peptides, proteins with apparent masses of 92, 90, 86, and 160 kDa were purified. Mass spectrometry data suggested that the 92- and 90-kDa polypeptides appear to be subtilisin-like proteins, whereas the 86-kDa polypeptide was identified as the molecular chaperone BiP of T. gondii.
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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Blotting, Western
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Carrier Proteins / analysis
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Carrier Proteins / chemistry
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Cells, Cultured
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Chlorocebus aethiops
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Chromatography, Affinity
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Endoplasmic Reticulum Chaperone BiP
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Fibroblasts / parasitology
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Fungal Proteins*
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Glutathione Transferase / genetics
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Glutathione Transferase / physiology
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Green Fluorescent Proteins
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Heat-Shock Proteins*
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Humans
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Indicators and Reagents / chemistry
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Luminescent Proteins / chemistry
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Mass Spectrometry
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Molecular Chaperones / analysis
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Molecular Chaperones / chemistry
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Molecular Sequence Data
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Organelles / chemistry*
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Peptide Mapping
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Protein Sorting Signals / physiology
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Ribosomal Proteins / chemistry*
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Ribosomal Proteins / genetics
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Toxoplasma / genetics
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Toxoplasma / physiology*
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Toxoplasma / ultrastructure
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Transfection
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Vero Cells / parasitology
Substances
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Carrier Proteins
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Endoplasmic Reticulum Chaperone BiP
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Fungal Proteins
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Heat-Shock Proteins
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Indicators and Reagents
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Luminescent Proteins
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Molecular Chaperones
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Protein Sorting Signals
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Ribosomal Proteins
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Su12 protein, Podospora anserina
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Green Fluorescent Proteins
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Glutathione Transferase