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Interaction of a novel cysteine and histidine‐rich cytoplasmic protein with galectin‐3 in a carbohydrate‐independent manner
Author(s) -
Me Rajesh P.,
Strom Molly,
Hughes R.Colin
Publication year - 2000
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(00)01310-7
Subject(s) - cytoplasm , complementary dna , galectin , biology , cysteine , cdna library , histidine , biochemistry , microbiology and biotechnology , open reading frame , peptide sequence , amino acid , gene , enzyme
We have used the yeast two‐hybrid system to search for cytoplasmic proteins that might assist in the intracellular trafficking of the soluble β‐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.