Preparatory Study On α-synuclein And γ-synuclein Structure And Function

Synuclein is a protein family having highly homologous .Synuclein family members are not found outside vertebrates. The synuclein family consists of three distinct genes, a-synuclein, P-synuclein, and y-synuclein.The proteins are found primarily in presynaptic terminals and karyotheca.Now, we know that two missense mutations (A53T and A30P) in a-synuclein was genetically linked to early-onset, familial Parkinson's disease, and it's the missense mutations in a-synuclein are not present in sporadic Parkinson's disease. Now, we do not have any strategy to research the functions of the wild a-synuclein (SNCA) and the two missense mutations. It is the best strategy that crystalling of protein and resolving the protein's structure, however, speculating that function in normal or abnormal, benign or malignant tissue. We obtained the a-synuclein (SNCA) cDNA by RT-PCR from fetus brain RNA, and constructed the prokaryotic expression vector, pGEX-4T-l-synuclein (SNCA) .Then ,we attained the crystal of the a-synuclein (SNCA) protein. Simutaneity, we cloned the missense mutation in a-synuclein A53T into pGEM-T vector.Now, people know little about -synuclein. We expected to find that it is concerned with any signal pathway, or cell framework or nuclear framework by finding the interactive protein. For the reason, we abtained the synuclein (SNCG) cDNA by RT-PCR from fetus brain RNA, and constructed the prokaryotic expression vector, pGEX-4T-l-synuclein (SNCG) , expression and purification the SNCGprotein, distilled the normal brain protein. We finded two interactive proteincrystallin alphaB and AHI-1 by GST-PULLDOWN and mass spectrum analysis. The first one is called heat shock 20KD like protein, it is the basic subunit of crystallin alpha protein. Crystallin alpha was first identified from vertebrate lens, now people finded that it existed in many tissue outside from lens, including the center nerve system. Crystallin alpha is a chaperone protein and belong to small heat shock protein family. Crystallin alphaB is a marker molecular of many neurodegeneration diseases. AHI1 is a noval gene.Now, we know nothing about this gene.synuclein (SNCG) have a difference in expression among diverse tissue and the benign or malignant tissue. In breast-cancer, the expression difference is regulated by DNA methylation of synuclein (SNCG) CpG island. We speculate that the expression difference of synuclein (SNCG) in normal brain and brain cancer is also regulated by DNA methylation of synuclein (SNCG) CpG island. Consequently, we distilled genome DNA from normal brain and brain cancer, and amplified the synuclein (SNCG) CpG island by PCR.