The Screening Of Proteins Interacting With MEF2C And CFL2 In Mouse Muscle Fibers

ObjectiveTo screen the proteins interacting with MEF2 C from mouse myoblasts C2C12 and CFL2 from mouse skeletal muscle in leg respectively,which could provide a theoretical foundation for further research of these two proteins on muscle development in mouse.MethodsInteraction protein compounds were extracted by co-immunoprecipitation,which were separated by SDS-PAGE and verified by Western Blotting.The differences strips were cut by enzyme trypsin.The peptides had been also successfully separated by liquid chromatography,and then analyzed identificated by electrospray Ionizsationmass.Cell protein profiling interacting with MEF2C and tissue protein profiling with CFL2 were obtained respectively.The screened proteins were retested by co-immunoprecipitation and Western Blotting.Results1.A differential band was successfully isolated by SDS-PAGE from total protein of C2C12 cells,which contained proteins of Beta-actin,GAPDH,LDHA and MDHM using mass spectrometry.The screened proteins were verified by Western Blotting and showed that these proteins were interacted with MEF2 C in the native state.2.The protein Ndufb7 interacting with CFL2 from mouse skeletal muscle inleg was verified by co-immunoprecipitation,mass spectrometry and Western Blotting in the native state.Conclusions1.Four proteins of ?-actin,GAPDH,LDHA and MDHM interacting with MEF2 C had been successfully screened and identified by co-immunoprecipitation and mass spectrometry analysis for the first time.2.The protein Ndufb7 had been also successfully achieved to interact with CFL2 by co-immunoprecipitation and mass spectrometry analysis firstly.