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Updated: Aug 14, 2026

Identifying Protein-protein Interaction Sites Using Peptide Arrays
Published on: November 18, 2014
Mapping of the interaction sites between apoptosis linked gene ALG-2 and HEED
1Bio-Med RRC and Division of Life Sciences, Pai Chai University, Taejon, Korea.
Abstract:
The apoptosis linked gene (ALG-2) is a 22 kDa Ca2+-binding protein of the penta EF-hand motif family. ALG-2 was discovered in a "death trap" assay using T-cell receptor-mediated apoptosis. Depletion of ALG-2 using an anti-sense ALG-2 construct inhibits apoptosis that is induced by several stimuli, such as staurosporin, dexamethason, Fas, and glucocorticoid. The Ca2+-dependent function of ALG-2 is consistent with the observation that the cytoplasmic Ca2+ level is elevated in apoptotic cells. We found that ALG-2 interacted specifically with the carboxy-terminal region of HEED using a yeast two-hybrid assay system. The mutants of HEED were constructed by deleting five WD repeat motifs one by one from the C-terminus of the protein. These mutants of ALG-2 were made by combining the EF hand Ca2+ binding motifs in various ways. Mapping of the interaction sites, using each of the mutants, revealed that the interaction between HEED and a third EF-hand motif of ALG-2 was stronger than the other combination.
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