Cloning, purification, crystallization and preliminary crystallographic analysis of acylphosphatase from Pyrococcus horikoshii OT3.
Acylphosphatase is one of the smallest enzymes and catalyzes the hydrolysis of the carboxy-phosphate bond. An extremely thermostable acylphosphatase from a hyperthermophilic archaea, Pyrococcus horikoshii OT3, has been cloned, expressed in Escherichia coli, purified and crystallized using the sitting-drop vapour-diffusion method with potassium/sodium tartrate as the precipitant at pH 5.5. X-ray diffraction data have been collected to a highest resolution of 1.72 angstroms on a synchrotron-radiation source. The crystals belong to space group P3(2)21, with approximate unit-cell parameters a = b = 86.6, c = 75.4 angstroms and two monomers in the asymmetric unit.
Miyazono
Ken Ichi
KI
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Kudo
Norio
N
Tanokura
Masaru
M
D017766:Acid Anhydride Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D002244:Carbon / Q000737:chemistry; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D004058:Diffusion; D004926:Escherichia coli / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D015233:Models, Statistical; D010710:Phosphates / Q000737:chemistry; D043902:Pyrococcus horikoshii / Q000201:enzymology*; D017356:Synchrotrons; D013644:Tartrates / Q000494:pharmacology; D014961:X-Ray Diffraction; D000097024:Acylphosphatase
D017766:Acid Anhydride Hydrolases; D002244:Carbon; D010710:Phosphates; D013644:Tartrates; D000097024:Acylphosphatase; C029768:tartaric acid