Author:

Keywords:

Science & Technology, Life Sciences & Biomedicine, Biochemical Research Methods, Biochemistry & Molecular Biology, Yeast two-hybrid, Two-hybrid array, Protein-protein interaction, GAL4, Yeast strain, PDZ interactions, PROTEIN INTERACTIONS, SYSTEMS, Protein–protein interaction, Humans, PDZ Domains, Protein Binding, Protein Interaction Mapping, Proteins, Saccharomyces cerevisiae, Two-Hybrid System Techniques, 0399 Other Chemical Sciences, 0601 Biochemistry and Cell Biology, Developmental Biology, 3101 Biochemistry and cell biology, 3404 Medicinal and biomolecular chemistry

Abstract:

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 KD in the high μmolar range) with any candidate protein among a panel of 266 individual clones, thereby comprehensively identifying its PDZ interactome.