caa a22 4500 397556225 CHVBK 20180308164803.0 cr unu---uuuuu 161202e199606 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021356 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021356 Crystallization of Expressed Porcine Kidney D-Amino Acid Oxidase and Preliminary X-Ray Crystallographic Characterization [Elektronische Daten] [Chiaki Setoyama, Retsu Miura, Yasuzo Nishina, Kiyoshi Shiga, Hisashi Mizutani, Ikuko Miyahara, Ken Hirotsu] The cDNA for porcine kidney D-amino acid oxidase (DAO) was cloned by means of the reverse transcription-polymerase chain reaction system from porcine kidney RNA and over-expressed in Escherichia coli which had been transformed with a vector containing the DAO cDNA. The expressed DAO was purified to homogeneity by a three-step procedure, i.e., heat-treatment, DEAE Sepharose column chromatography, and hydroxyapatite column chromatography. The purified DAO preparation, rDAO (recombinant DAO), showed an identical UV-visible absorption spectrum and catalytic activity with those of the wild-type enzyme purified from porcine kidney. Crystallization of rDAO was performed by the hanging-drop method and crystals of suitable quality for X-ray crystallography were obtained. The crystals so obtained diffracted to 2.5 Å with a conventional X-ray source, and to 2.0 Å with synchrotron radiation. The crystals belong to the orthorhombic space group P212121 with unitcell dimensions of a = 110.3, b = 92.9, c=71.6 Å. A Vm value of 2.35 A3/Da indicates that there are two subunits related by a twofold non-crystallographic axis in the asymmetric unit. Two heavy atom derivatives have been identified. © by the Journal of Biochemistry Regular Papers nationallicence crystallization nationallicence D-amino acid oxidase nationallicence expression nationallicence flavoenzyme nationallicence X-ray crystallography nationallicence Setoyama Chiaki Departments of Biochemistry Honjo, Kumamoto 860 aut Miura Retsu Departments of Biochemistry Honjo, Kumamoto 860 aut Nishina Yasuzo Physiology, Kumamoto University School of Medicine Honjo, Kumamoto 860 aut Shiga Kiyoshi Physiology, Kumamoto University School of Medicine Honjo, Kumamoto 860 aut Mizutani Hisashi Department of Chemistry, Faculty of Science, Osaka City University Sugimoto, Sumiyoshi-ku, Osaka 558 aut Miyahara Ikuko Department of Chemistry, Faculty of Science, Osaka City University Sugimoto, Sumiyoshi-ku, Osaka 558 aut Hirotsu Ken Department of Chemistry, Faculty of Science, Osaka City University Sugimoto, Sumiyoshi-ku, Osaka 558 aut The Journal of Biochemistry Oxford University Press 119/6(1996-06), 1114-1117 0021-924X 119:6<1114 1996 119 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021356 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021356 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Setoyama Chiaki Departments of Biochemistry Honjo, Kumamoto 860 aut NATIONALLICENCE 700 1- Miura Retsu Departments of Biochemistry Honjo, Kumamoto 860 aut NATIONALLICENCE 700 1- Nishina Yasuzo Physiology, Kumamoto University School of Medicine Honjo, Kumamoto 860 aut NATIONALLICENCE 700 1- Shiga Kiyoshi Physiology, Kumamoto University School of Medicine Honjo, Kumamoto 860 aut NATIONALLICENCE 700 1- Mizutani Hisashi Department of Chemistry, Faculty of Science, Osaka City University Sugimoto, Sumiyoshi-ku, Osaka 558 aut NATIONALLICENCE 700 1- Miyahara Ikuko Department of Chemistry, Faculty of Science, Osaka City University Sugimoto, Sumiyoshi-ku, Osaka 558 aut NATIONALLICENCE 700 1- Hirotsu Ken Department of Chemistry, Faculty of Science, Osaka City University Sugimoto, Sumiyoshi-ku, Osaka 558 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 119/6(1996-06), 1114-1117 0021-924X 119:6<1114 1996 119 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford