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Imaging-plate Detector (imaging-plate + detector)
Selected AbstractsAway from the edge II: in-house Se-SAS phasing with chromium radiationACTA CRYSTALLOGRAPHICA SECTION D, Issue 7 2005Hao Xu Recently, the demands of high-throughput macromolecular crystallography have driven continuous improvements in phasing methods, data-collection protocols and many other technologies. Single-wavelength anomalous scattering (SAS) phasing with chromium X-ray radiation opens a new possibility for phasing a protein with data collected in-house and has led to several successful examples of de novo structure solution using only weak anomalous scatterers such as sulfur. To further reduce data-collection time and make SAS phasing more robust, it is natural to combine selenomethionine-derivatized protein (SeMet protein) with Cr,K, radiation to take advantage of the larger anomalous scattering signal from selenium ( = 2.28 e,) compared with sulfur ( = 1.14 e,). As reported herein, the crystal structure of a putative chorismate mutase from Clostridium thermocellum was determined using Se-SAS with Cr,K, radiation. Each protein molecule contains eight selenomethionine residues in 148 amino-acid residues, providing a calculated Bijvoet ratio of about 3.5% at the Cr,K, wavelength. A single data set to 2.2,Å resolution with approximately ninefold redundancy was collected using an imaging-plate detector coupled with a Cr source. Structure solution, refinement and deposition to the Protein Data Bank were performed within 9,h of the availability of the scaled diffraction data. The procedure used here is applicable to many other proteins and promises to become a routine pathway for in-house high-throughput crystallography. [source] Structural studies of MIP synthaseACTA CRYSTALLOGRAPHICA SECTION D, Issue 3 2000Adam J. Stein The conversion of glucose 6-phosphate to 1- l - myo -inositol 1-phosphate (MIP) by 1- l - myo -inositol 1-phosphate synthase (MIP synthase) is the first committed and rate-limiting step in the de novo biosynthesis of inositol in all eukaryotes. The importance of inositol-containing molecules both as membrane components and as critical second messenger signal-transduction species make the function and regulation of this enzyme important for a host of biologically important cellular functions including proliferation, neurostimulation, secretion and contraction. MIP synthase has been overexpressed in Esherichia coli and purified to homogeneity by chromatographic methods. Two crystal forms of MIP synthase were obtained by the hanging-drop vapor-diffusion method. Native data sets for both crystal forms were collected in-house on a Rigaku R-AXIS IIC imaging-plate detector. Crystal form I belongs to space group C2, with unit-cell parameters a = 153.0, b = 96.6, c = 122.6,Å, , = 126.4°, and diffracts to 2.5,Å resolution. Crystal form II belongs to space group P21, with unit-cell parameters a = 94.5, b = 186.2, c = 86.5,Å, , = 110.5°, and diffracts to 2.9,Å resolution. [source] Crystallization of the pneumococcal autolysin LytC: in-house phasing using novel lanthanide complexesACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 4 2010Inmaculada Pérez-Dorado LytC, one of the major autolysins from the human pathogen Streptococcus pneumoniae, has been crystallized as needles by the hanging-drop technique using 10%(w/v) PEG 3350 as precipitant and 10,mM HEPES pH 7.5. LytC crystals were quickly soaked in mother liquor containing 2,mM of the complex Gd-HPDO3A to produce derivatized crystals (LytCGd-HPDO3A). Both native LytC and isomorphous LytCGd-HPDO3A crystals were flash-cooled in a nitrogen flow at 120,K prior to X-ray data collection using an in-house Enraf,Nonius rotating-anode generator (, = 1.5418,Å) and a MAR345 imaging-plate detector. In both cases, good-quality diffraction patterns were obtained at high resolution. LytCGd-HPDO3A crystals allowed the collection of a SAD X-ray data set to 2.6,Å resolution indexed in terms of a P21 monoclinic unit cell with parameters a = 59.37, b = 67.16, c = 78.85,Å, , = 105.69°. The anomalous Patterson map allowed the identification of one heavy-atom binding site, which was sufficient for the calculation of an interpretable anomalous map at 2.6,Å resolution. [source] Improvement of SAXS measurements on Kratky slit systems by Göbel mirrors and imaging-plate detectorsJOURNAL OF APPLIED CRYSTALLOGRAPHY, Issue 3-2 2000Alexander Bergmann Laboratory X-ray sources emit a highly divergent beam. The Kratky compact camera is constructed to maximize the intensity in the sample using a slit collimation system. The performance of this camera can be further increased if the primary beam is collimated from a divergent into a parallel beam. A recently developed device for this purpose is the so-called `Göbel mirror'. This mirror is made of parabolically bent multilayers, designed to collimate divergent X-rays from laboratory X-ray sources into a parallel and monochromatic beam of high brilliance. Modification of the block collimation system in combination with a Göbel mirror leads to a different beam geometry, resulting in an intensity increase by a factor of about 10. The gain in intensity implicates the use of imaging-plate detectors, which have a wide linear range in intensity and allow the full use of the increased intensity. Hence the quality of the SAXS data is improved by the higher intensity primary beam, the much lower background due to the exclusive use of Cu K, radiation, and a detection unit which is linear in the measured intensity regime. All these advantages, such as intensity gain, lower background, better quality of the data, are demonstrated with some selected experimental results. [source] | ||||||||||