PURIFICATION AND CHARACTERIZATION OF FETAL SPECIFIC SYNAPTOSOMAL 50KDa PROTEIN

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In order to study changes of CNS functions and disorders by aging, we tried to find qualitative and quantitative changes of synaptosomal proteins by aging. The fetal specific 50K protein was purified from lyophilisate of ultracentrifugation sup of rat P_2 fraction after hypotonic and ultrasonic treatment.The fetal 50K protein was separated on preparative SDS-PAGE and extracted from gels into 0.1% SDS solution. The extract was further applied on Sephadex G-75 column pre-equilibrated with 20 mM imidazole-HC1 pH 7.0 containing 0.1% SDS. The fetal 50K protein was recovered after void volume fraction. At this step, the 50K protein was purified as a single band on SDS-PAGE. The partial amino acid sequence of one peptide fragment obtained by HPLC after trypsin digestion was as follow ; N-F(A)ILVFLE(A)PFNMDFVW-. This sequence showed partial homology with polyoma viral DNA binding protein by computer search. When the fetal 50K was added in culture medium of neurobalstoma cells, ^3H-thymidine uptake was increased 2 times at 10^<-8>M.