Expression,Purification and Structural Characterization of Mouse Talin-1 Head Δ139-168

Talin-1 head (hereinafter referred to as TH) is the head structure of talin protein,which contains a four-point-one-protein/ezrin/radixin/moesin (FERM) domain.Its F1 domain contains an unstructured loop of 30 amino acids (139-168),which does not interact with other domains.Because TH doesn't get the crystal structure and whether the unstructured loop has obvious influence on the TH secondary structure,therefore,the truncated talin-1 head Δ139-168 (hereinafter referred to as THΔ) was constructed and its structure and the impact of stability after truncation were analyzed.Molecular biology and structural biology methods were used to construct prokaryotic expression vectors of TH and THΔ,explore and optimize the expression conditions of recombinants,and they were purified by affinity chromatography and FPLC gel filtration chromatography.Finally,a large number of stable,high-purity protein samples were prepared successfully.The physicochemical properties and structural stability of the proteins were investigated by dynamic light scattering and circular dichroism.The results showed that the THΔ secondary structure of the truncated body did not change significantly,the structural stability was enhanced and the resistance to guanidine hydrochloride and high temperature was stronger.