The aim of this work is to investigate specific features of α-synuclein synthesis in eukaryotic cells compared to prokaryotic ones, namely to test the possible formation of Tyr136Cys mutant protein in eukaryotic cells. It has previously been shown that Tyr136Cys substitution occurs upon expression of the wild-type α-synuclein gene in E. coli cells, since UAC triplet encoding Tyr136 is erroneously recognized by the bacterial translational system as UGC cysteine triplet. We obtained SH-SY5Y human neuroblastoma cell lines producing wild-type α-synuclein (transiently transfected as well as stable lines). The production of α-synuclein in different neuroblastoma lines was confirmed by immunofluorescence analysis. The presence of different forms of α-synuclein was determined by immunoblotting after SDS-PAGE under non-reducing conditions. This approach allows to detect Tyr136Cys α-synuclein by the emergence of α-synuclein dimers due to crosslinking of the monomers by disulfide bonds. It was shown that the expression of α-synuclein in eukaryotic cells does not lead to the formation of the dimeric forms of the protein that were observed during its expression in the bacterial system. Thus, Tyr136Cys substitutions are typical for the expression of α-synuclein in bacteria, but not in eukaryotes.