Development of Improved Methods for Protein Separation and Identification

For identification of proteins, able to connect specifically or non-specifically with peptide SCGN, protein material from RIN-5F rat insulinoma malignant cells and E. coli bacteria strains, both with inserted by transfection with recombinant vectors rat gene SCGN, was isolated. Subsequently, SCGN peptide was isolated, precipitated and incubated with lysates from rat pancreas and brain, known as anatomic organs with highest expression of SCGN gene. For determination of proteins with highest affinity to this target peptide, from both organs, it was separately mixed with both lysates, and the mixtures were subjected on separation on CNBr-Sepharose and GST-Agarose columns, respectively. GST-Agarose technique indicated some advantages in comparison with CNBr-Sepharose, connected mainly with higher yields of different protein types because of the escaped protein degradation influence of CNBr. The results supported the suggested in our previous studies abilities of protein SCGN to connect with cytoskeleton elements, in confirmation with some literature findings. On this base, a mechanism for indirect influence of this peptide on the control of cell growth and proliferation, has been proposed. Future studies are necessary in this direction