Crystallographic analysis of the FACT-histone complex

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FACT, which interacts with the histone H2A-H2B complex, is known to be involved in nucleosome assembly/disassembly. In order to reveal the molecular mechanism of nucleosome assembly/disassembly, crystal structure analysis of FACT was initiated. In the present study, we have established a method that can give about 7 mg of highly purified FACT from 1L culture of insect cell SF+. In addition, a purification method of histone proteins was improved. We confirmed that the purified FACT interacts with the histone proteins. To establish a proper condition for preparing the FACT-histone complex without forming aggregates, solution conditions of the FACT-histone complex were investigated by the dynamic light scattering method. On the basis these results, we are making an effort to obtain crystals of the FACT-histone complex.