Tim Heacock Identification of protein interactions of the intracellular domain of Neurotactin by biopanning of a phage display cDNA library Division Sciences Department Biology Year 2004 Advisor Eric Liebl

Neurotactin (Nrt) is a transmembrane cell adhesion molecule that is involved in axon guidance during Drosophila melanogater development.  The extracellular domain of Nrt interacts with the secreted protein Amalgam to promote cell:cell adhesion. The interactions of the intracellular domain of Nrt (Nrt-IC), however, are uncharacterized.  We attempted to identify the protein:protein interactions of  Nrt-IC.  To accomplish this goal, we constructed a T7 phage display cDNA library using D. melanogaster embryo mRNA primed with HindIII random primers.  Nrt-IC interacting phage were selected for by biopanning over purified Nrt-IC.  The screen identified Nrt-IC interactions with phage that encode the neuronally expressed Bangles and beads (Bnb) and the kinase domain of CG1344.  However, specific affinity of the phage that encode these cDNAs could not be demonstrated.  Future experiments to confirm these interactions include finding conditions where specific affinity for Nrt-IC can be identified and testing for genetic interactions between bnb, CG1344, and nrt.