E.coli'de tam boyutlu ve kesilmiş yeni CCDC-124 proteinin klonlaması, ekspresyonu ve biyokimyasal özellikleri

Coiled-coil domain-containing 124 (CCDC-124) protein is an eukaryotic protein that is conserved with a spectrum extends low to high eukaryotes. Despite CCDC-124 mainly localizes centrosome and midbody with a correlation its cytokinetic role, it also seems to be as a shuttle between nuclear and cytoplasmic complexes. CCDC-124 contains at least one putative RNA-binding domain. According to our previous studies of mass spectrometry-based proteomics and their validations CCDC-124 appears to be related to nuclear and cytoplasmic phase-separated complexes. To date many studies have been done to identify the function and properties of the CCDC-124 protein. In this study, we aimed to investigate the functional role of CCDC-124 protein fragments on biochemical and biophysical properties. We have prepared four truncated mutants: one without a C-terminus and one without N-terminus required for the 32 kDa isoform, the other two mutants are designed according to the putative RNA-binding region. To determine the physicochemical properties of full length and truncated fragments of CCDC-124 protein, we expressed and purified the protein form E. coli cells. Then purified proteins subjected to various chemical agents. Thus we could try to understand if the proteins have phase separation behavior. Our results provide useful information about the physiological role of the protein.