Structure Researchs On Proteus Mirabilis Cap Protein FliD And Human Chromosomal Passenger Complex

This thesis contains two parts: preliminary X-ray researchs on proteusmirabilis cap protein FliD and human chromosomal passenger complex.Bacterial flagella, Directively participate in pathogen infection. Theflagellum is composed of about50varieties of proteins which can be dividedinto three parts, including the basal body (a rotary motor region), thefilament (a helical propeller region) and the hook (a universial joint linkingthe motor and propeller). The number and length of flagella are closelyassociated with flagella cap protein FliD, but for the flagellum assemblymechanism is not clear. This study we obtained a lot of proteus mirabilis capprotein FliD by cloning, expression and purification method. After obtainedthe protein crystals, we make its diffraction ability from no to3.5throughlimiting protease hydrolysis experiment and crystal dehydration.Additionally, structural biology reaearch about about complexes of FliD andthe chaperone FliT are underway.The kinase Aurora B, together with INCENP, Borealin andSurvivin,form the chromosomal passenger complex (CPC), which plays multiple roles during mitosis and meiosis by changing its localization. Apurified ternary complex of Survivin, Borealin-INCENP (1-58aa), speciallybound to H3T3ph petide in vitro, Survivin also was able to bind to H3T3phpetides in the absence of other CPC subunits by nuclear magnetic resonance(NMR) spectroscopy, implying that it is the receptor for H3T3ph. Weobtained a series of subcomplex by cloning, expressing and purification.Whereas, the stability of Survivin expression alone is bad, and there is adegradation of stripe. But with Borealin (10-109aa) building a subcomplex isthe degradation stripes disappear. Finally we choose cloning, expression andpurification Survivin-Borealin (10-109aa) in vitro, and obtained a gooddiffraction ability crystal with H3T3ph by biosynthesis. But the electrondensity of the H3T3ph combination has not been found in crystal structure,and the flollow-up studies of the complex are carried out continually.