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PMID:10745073
Citation |
Menon, RP, Strom, M and Hughes, RC (2000) Interaction of a novel cysteine and histidine-rich cytoplasmic protein with galectin-3 in a carbohydrate-independent manner. FEBS Lett. 470:227-31 |
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Abstract |
We have used the yeast two-hybrid system to search for cytoplasmic proteins that might assist in the intracellular trafficking of the soluble beta-galactoside-binding protein, galectin-3. We utilised as bait murine full-length galectin-3 to screen a murine 3T3 cDNA library. Several interacting clones were found to encode a partial open reading frame and a full-length clone was obtained by rapid amplification of cDNA ends methodology. In various assays in vitro the novel protein was shown to bind galectin-3 in a carbohydrate-independent manner. The novel protein contains an unusually high content of cysteine and histidine residues and shows significant sequence homologies with several metal ion-binding motifs present in known proteins. Confocal immunofluorescence microscopy of permeabilised 3T3 cells shows a prominent perinuclear, as well as cytoplasmic, localisation of the novel protein. |
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Keywords |
3T3 Cells; Amino Acid Motifs; Amino Acid Sequence; Animals; Antigens, Differentiation/chemistry; Antigens, Differentiation/genetics; Antigens, Differentiation/metabolism; Binding Sites; Carbohydrate Metabolism; Cell Nucleus/chemistry; Cloning, Molecular; Conserved Sequence/genetics; Cysteine/genetics; Cysteine/metabolism; Cytoplasm/chemistry; Galectin 3; Histidine/genetics; Histidine/metabolism; Humans; Mice; Microscopy, Fluorescence; Molecular Sequence Data; Peptide Fragments/chemistry; Peptide Fragments/genetics; Peptide Fragments/metabolism; Precipitin Tests; Protein Binding; Proteins/chemistry; Proteins/genetics; Proteins/metabolism; RNA, Messenger/analysis; RNA, Messenger/genetics; Sequence Alignment; Two-Hybrid System Techniques |
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Annotations
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