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MAD Phasing Method (mad + phasing_method)
Selected AbstractsCrystallization and preliminary X-ray analysis of the archaeosine tRNA-guanine transglycosylase from Pyrococcus horikoshiiACTA CRYSTALLOGRAPHICA SECTION D, Issue 11 2001Ryuichiro Ishitani The archaeosine tRNA-guanine transglycosylase from the hyperthermophilic archaeon Pyrococcus horikoshii was crystallized and preliminary X-ray characterization was performed. Single crystals were grown by the hanging-drop vapour-diffusion method, using sodium/potassium phosphate and sodium acetate as precipitants. The space group is P41212 or P43212, with unit-cell parameters a = b = 99.28,(14), c = 363.74,(56),Å. The cryocooled crystals diffracted X-rays beyond 2.2,Å resolution using synchrotron radiation from station BL44XU at SPring-8 (Harima). Selenomethionine-substituted protein crystals were prepared in order to solve the structure by the MAD phasing method. [source] Expression, crystallization and preliminary crystallographic studies of a novel bifunctional N -acetylglutamate synthase/kinase from Xanthomonas campestris homologous to vertebrate N -acetylglutamate synthaseACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 12 2006Dashuang Shi A novel N -acetylglutamate synthase/kinase bifunctional enzyme of arginine biosynthesis that was homologous to vertebrate N -acetylglutamate synthases was identified in Xanthomonas campestris. The protein was overexpressed, purified and crystallized. The crystals belong to the hexagonal space group P6222, with unit-cell parameters a = b = 134.60, c = 192.11,Å, and diffract to about 3.0,Å resolution. Selenomethionine-substituted recombinant protein was produced and selenomethionine substitution was verified by mass spectroscopy. Multiple anomalous dispersion (MAD) data were collected at three wavelengths at SER-CAT, Advanced Photon Source, Argonne National Laboratory. Structure determination is under way using the MAD phasing method. [source] Crystallization and preliminary X-ray crystallographic analysis of the catalytic domain of pyrrolysyl-tRNA synthetase from the methanogenic archaeon Methanosarcina mazeiACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 10 2006Ryohei Ishii Pyrrolysyl-tRNA synthetase (PylRS) from Methanosarcina mazei was overexpressed in an N-terminally truncated form PylRS(c270) in Escherichia coli, purified to homogeneity and crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol as a precipitant. The native PylRS(c270) crystals in complex with an ATP analogue belonged to space group P64, with unit-cell parameters a = b = 104.88, c = 70.43,Å, , = , = 90, , = 120°, and diffracted to 1.9,Å resolution. The asymmetric unit contains one molecule of PylRS(c270). Selenomethionine-substituted protein crystals were prepared in order to solve the structure by the MAD phasing method. [source] Expression, purification, crystallization and preliminary X-ray crystallographic studies of a novel acetylcitrulline deacetylase from Xanthomonas campestrisACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 7 2005Dashuang Shi A novel N -acetyl- l -citrulline deacetylase that is able to catalyze the hydrolysis of N -acetyl- l -citrulline to acetate and citrulline was identified from Xanthomonas campestris. The protein was overexpressed, purified and crystallized. The crystals belong to the monoclinic space group C2 and diffract to 1.75,Å resolution, with unit-cell parameters a = 94.13, b = 95.23, c = 43.61,Å, , = 93.76°. Since attempts to use homologous structural models to solve the structure via molecular replacement were unsuccessful, the selenomethionine-substituted protein was prepared using an overnight auto-induction overexpression system. Selenomethionine incorporation into the protein was verified by MALDI,TOF/TOF mass-spectroscopic analysis after trypsin digestion. The crystals of the selenomethionine-substituted protein were prepared using crystallization conditions similar to those for the native protein. Multiple anomalous dispersion (MAD) data were collected at Brookhaven National Laboratory. Structure determination is under way using the MAD phasing method. [source] | ||||||||||