Crystallization And Preliminary X-ray Analysis Of Spindlin1 & Screening The Putative Partner Peptides Of Spindlin1

Spindlinl is a protein of 237 amino-acid residues which is a member of Spin/Ssty family. Its function and structural bases remain largely unknown. In this study, Spindlinl was expressed as GST-fusion protein in Escherichia coli and the GST-Spindlinl was soluble. We harvested the highly-purified Spindlinl by affinity chromatography and ani on-exchange chromatography. The recombinant Spindlinl was crystallized using the hanging-drop vapour-diffusion method, which yielded needle-shaped crystals. After optimized the crystallization conditions, we obtained diffraction-quality crystals. Finally the crystal structure of Spindlin was built and refined, and the result had been submitted to PDB.Based on the bioinformatics analysis and the crystal structure, the site-directed mutagenesis was used and 18 mutant vectors were gained. We transfected the vectors into NIH 3T3 cell. Further assays would be performed.