Selection of ligands by panning of domain libraries displayed on phage lambda reveals new potential partners of synaptojanin 1.

摘要: One of the goals functional genomics is description reliable and complete protein interaction networks. To facilitate ligand discovery from complex mixtures, we have developed an improved approach that affected by a negligible fraction false positives. We combined novel technique based on display cDNA libraries capsid bacteriophage lambda efficient plaque assay to reveal phage displaying ligands are enriched after only couple affinity purification steps. show system has unique ability display, at high density, proteins ranging in size few least 300 amino acid residues. This characteristic permits attenuation bias selection procedure and, same time, offers sensitive us do away with background without unduly increasing number cycles. By using proline-rich fragment synaptojanin 1 as bait, identified, brain library, seven all containing either SH3 or WW domains. Four these correspond already been validated physiological partners, while remaining three new whose relevance remains be established. Two different regions p21-activated kinase (Pak1) WAVE/SCAR2 retrieve library