The Mechanism Study Of Aggregation Of EWSR1 Protein Prion-like Domains Associated With Neurodegenerative Disease

Ewing Sarcoma(EWS)is the second most common cartilage malignancy in adolescents.Patients suffer great pain and have low survival rates.Ewing sarcoma breakpoint region 1(EWSR1),which contains prion protein domains(PrDs)rich in glutamine,is the main cause of the disease,Studies on Parkinson's disease(PD)revealed similar repetitive poly-glutamine regions in a-synuclein,which is critical for the aggregation and pathogenesis of the disease.In this work,We synthesized peptide segments and expression of longer protein fragments of the PrLDs of EWSR1 and explored their aggregation in order to find the potential connection between amyloid formation and poly-glutamine patterns.Therefore,We can rise some possible mechanism of the pathogenesis of neurodegenerative diseases at the molecular level.Herein,the peptide fragments in the PrLDs region were synthesized chemically.The secondary structures of these peptides before and after the aggregation were examined by means of elemental dichroism(CD).The fluorescent dye thioflavin T(ThT)was used to monitor the kinetic parameters of the peptide aggregation in real time.Transmission electron microscopy(TEM)and atomic force microscopy(AFM)were used to characterize the morphology of the fibers formed by the aggregation.We have observed that all three fragments aggregate into fibers,and obtained the kinetic curve of fiber growth,recording the various stages of aggregation.Additionally,we expressed the longer fragment EWSR1(1-280)and studied its aggregation behavior.In sum,we have found key segments for EWSR1 aggregation,providing insights for mechanism of the pathogenesis and potential targets for the inhibitor development.