Identification of PDZ Interactions by Yeast Two-Hybrid Technique

The yeast two-hybrid technique is a powerful method to detect direct protein-protein interactions. Due to its accessibility, speed, and versatility, this technique is easy to set up in any laboratory and suitable for small and large scale screenings. Here we describe the implementation of an array-based screening that allows for the probing of the entire human PDZ ORFeome (or hPDZome) by yeast two-hybrid technique. With this approach, one can rapidly identify the PDZ domains that are able to interact (up to K_D in the high μmolar range) with any candidate protein among a panel of 266 individual clones, thereby comprehensively identifying its PDZ interactome.