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Grease matrix as a versatile carrier of proteins for serial crystallography
Sugahara, Michihiro,Mizohata, Eiichi,Nango, Eriko,Suzuki, Mamoru,Tanaka, Tomoyuki,Masuda, Tetsuya,Tanaka, Rie,Shimamura, Tatsuro,Tanaka, Yoshiki,Suno, Chiyo,Ihara, Kentaro,Pan, Dongqing,Kakinouchi, Ke Nature Publishing Group 2015 NATURE METHODS Vol. No.
Serial femtosecond X-ray crystallography (SFX) has revolutionized atomic-resolution structural investigation by expanding applicability to micrometer-sized protein crystals, even at room temperature, and by enabling dynamics studies. However, reliable crystal-carrying media for SFX are lacking. Here we introduce a grease-matrix carrier for protein microcrystals and obtain the structures of lysozyme, glucose isomerase, thaumatin and fatty acid–binding protein type 3 under ambient conditions at a resolution of or finer than 2 Å.
( Sung Bin Park ),( Takashi Fujimoto ),( Eiichi Mizohata ),( Tsuyoshi Inoue ),( Moon Hee Sung ),( Hiroshi Uyama ) 한국미생물 · 생명공학회 2013 Journal of microbiology and biotechnology Vol.23 No.7
Monoliths are functional porous materials with a three-dimensional continuous interconnected pore structure in a single piece. A monolith with uniform shape based on poly(γ-glutamic acid) (PGA) has been prepared via a thermally induced phase separation technique using a mixture of dimethyl sulfoxide, water, and ethanol as solvent. The morphology of the obtained monolith was observed by scanning electron microscopy and the surface area of the monolith was evaluated by the Brunauer Emmett Teller method. The effects of fabrication parameters such as the concentration and molecular mass of PGA and the solvent composition have been systematically investigated. The PGA monolith was cross-linked with hexamethylene diisocyanate to produce the water-insoluble monolith. The addition of sodium chloride to the phase separation solvent affected the properties of the cross-linked monolith. The swelling ratio of the cross-linked monolith toward aqueous solutions depended on the buffer pH as well as the monolith fabrication condition. Copper(II) ion was efficiently adsorbed on the cross-linked PGA monolith, and the obtained copper-immobilized monolith showed strong antibacterial activity for Escherichia coli. By combination of the characteristic properties of PGA (e.g., high biocompatibility and biodegradability) and the unique features of monoliths (e.g., through-pore structure, large surface area, and high porosity with small pore size), the PGA monolith possesses large potentials for various industrial applications in the biomedical, environmental, analytical, and separation fields.