Non-erythroid beta spectrin interacting proteins and their effects on spectrin tetramerization.

With yeast two-hybrid methods, we used a C-terminal fragment (residues 1697-2145) of non-erythroid beta spectrin (βII-C), including the region involved in the association with alpha spectrin to form tetramers, as the bait to screen a human brain cDNA library to identify proteins interacting with βII-C. We applied stringent selection steps ...
to eliminate false positives and identified 17 proteins that interacted with βII-C (IP(βII-C) s). The proteins include a fragment (residues 38-284) of "THAP domain containing, apoptosis associated protein 3, isoform CRA g", "glioma tumor suppressor candidate region gene 2" (residues 1-478), a fragment (residues 74-442) of septin 8 isoform c, a fragment (residues 704-953) of "coatomer protein complex, subunit beta 1, a fragment (residues 146-614) of zinc-finger protein 251, and a fragment (residues 284-435) of syntaxin binding protein 1. We used yeast three-hybrid system to determine the effects of these βII-C interacting proteins as well as of 7 proteins previously identified to interact with the tetramerization region of non-erythroid alpha spectrin (IP(αII-N) s) [1] on spectrin tetramer formation. The results showed that 3 IP(βII-C) s were able to bind βII-C even in the presence of αII-N, and 4 IP(αII-N) s were able to bind αII-N in the presence of βII-C. We also found that the syntaxin binding protein 1 fragment abolished αII-N and βII-C interaction, suggesting that this protein may inhibit or regulate non-erythroid spectrin tetramer formation.
Mesh Terms:
Binding Sites, Brain, Carrier Proteins, Cell Line, Cytoskeleton, Gene Library, Humans, Microfilament Proteins, Models, Molecular, Neurons, Peptide Fragments, Plasmids, Polymerization, Protein Binding, Protein Isoforms, Protein Structure, Tertiary, Recombinant Proteins, Transfection, Two-Hybrid System Techniques
Cell. Mol. Biol. Lett.
Date: Dec. 01, 2011
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