Screening of Hsp105alpha-binding proteins using yeast and bacterial two-hybrid systems.

Hsp105alpha is a 105-kDa stress protein, which is expressed constitutively at especially high levels in the brain compared with other tissues in mammals, and is also induced by a variety of stressors. Recently, we have shown that Hsp105alpha binds to alpha-tubulin and prevents the heat-induced disaggregation of microtubules. To further ...
elucidate the function of Hsp105alpha, we searched for Hsp105alpha-binding proteins by screening a mouse FM3A cell library and human and mouse brain cDNA libraries using the yeast and bacterial two-hybrid systems. We showed here that Hsp105alpha interacted with several cellular proteins, such as cofilin, dynein light chain 2A, alpha-adducin, ubiquitin activating enzyme E1, phosphoglycerate kinase 1, and platelet-activating factor acethylhydrolase alpha1-subunit. The interaction was validated by the results of a pull-down assay and indirect immunofluorescence analysis. The significance of Hsp105alpha and Hsp105alpha-binding proteins in cells was discussed.
Mesh Terms:
Animals, Bacterial Proteins, Brain, COS Cells, Calmodulin-Binding Proteins, Cytoskeleton, DNA, Complementary, Fluorescent Antibody Technique, Indirect, Fungal Proteins, Gene Library, HSP110 Heat-Shock Proteins, HSP70 Heat-Shock Proteins, Humans, Mice, Microscopy, Fluorescence, Microtubules, Plasmids, Protein Binding, Protein Biosynthesis, Recombinant Proteins, Transcription, Genetic, Tubulin, Two-Hybrid System Techniques
Biochem. Biophys. Res. Commun.
Date: Feb. 06, 2004
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