Purification and preliminary X-ray crystallographic studies of recombinant 7,8-diaminopelargonic acid synthase from Escherichia coli

1998 ◽  
Vol 54 (6) ◽  
pp. 1397-1398 ◽  
Author(s):  
Helena Käck ◽  
Katharine J. Gibson ◽  
Anthony A. Gatenby ◽  
Gunter Schneider ◽  
Ylva Lindqvist
Keyword(s):  
Escherichia Coli ◽  
Polyethylene Glycol ◽  
Pyridoxal Phosphate ◽  
Crystal Forms ◽  
X Ray ◽  
Space Groups ◽  
Cell Dimensions

Recombinant 7,8-diaminopelargonic acid synthase from Escherichia coli, a pyridoxal-phosphate-dependent aminotransferase, has been crystallized in space groups P21 and C2. Both crystal forms were obtained at pH 7.3 with 21% polyethylene glycol and 10% 2-propanol as precipitants. The cell dimensions were a = 130, b = 57.5, c = 117 Å, β = 110° for the C2 crystals, and a = 58.4, b = 55.6, c = 121 Å, β = 96.9° for the P21 crystals, which diffract to at least 2.6 and 2.0 Å resolution, respectively.

Crystallization and preliminary X-ray diffraction data of the complex of recombinant tick anticoagulant peptide (rTAP) and bovine factor Xa

1999 ◽  
Vol 55 (4) ◽  
pp. 862-864 ◽  
Author(s):  
Anzhi Wei ◽  
Angela Smallwood ◽  
Richard S. Alexander ◽  
Jodie Duke ◽  
Harold Ross ◽  
...  
Keyword(s):  
Polyethylene Glycol ◽  
Factor Xa ◽  
Asymmetric Unit ◽  
X Ray Diffraction ◽  
Crystal Forms ◽  
X Ray ◽  
Cell Dimensions ◽  
Unit Cell Dimensions ◽  
Tick Anticoagulant Peptide ◽  
Bovine Factor

The complex of bovine factor Xa and recombinant tick anticoagulant peptide (rTAP) was crystallized in two different crystal forms using polyethylene glycol as a precipitant. Form I belongs to space group P42212 with unit-cell dimensions a = b = 133.1, c = 68.8 Å. It contains one complex per asymmetric unit and diffracts to 3.0 Å resolution. Form II belongs to P41212 (or P43212) with dimensions a = b = 126.5, c = 146.7 Å; it contains two complexes per asymmetric unit and diffracts to 2.5 Å. The crystals of both forms consist of factor Xa (MW  = 45.3 kDa) and rTAP (MW = 6.7 kDa).

Preliminary crystallographic studies on glutamine synthetase from Mycobacterium tuberculosis

1999 ◽  
Vol 55 (4) ◽  
pp. 865-868 ◽  
Author(s):  
Harindarpal S. Gill ◽  
Gaston M.U. Pfluegl ◽  
David Eisenberg
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Glutamine Synthetase ◽  
Etiologic Agent ◽  
Asymmetric Unit ◽  
Crystal Forms ◽  
X Ray ◽  
Cell Dimensions ◽  
The One ◽  
Unit Cell Dimensions ◽  
Axes Of Symmetry

The etiologic agent of tuberculosis, Mycobacterium tuberculosis, has been shown to secrete the enzyme glutamine synthetase (TB-GS) which is apparently essential for infection. Four crystal forms of a recombinant TB-GS were grown. The one chosen for synchrotron X-ray data collection belongs to space group P212121 with unit-cell dimensions 208 × 258 × 274 Å, yielding 2.4 Å resolution data. A Matthews number of 2.89 Å3 Da−1 is found, corresponding to 24 subunits of molecular mass 1300 kDa in the asymmetric unit. From earlier work, the structure of Salmonella typhimurium GS, which is 51% identical in sequence to TB-GS, is known to be dodecameric with 622 symmetry. Self-rotation calculations on the TB-GS X-ray data reveal only one set of sixfold and twofold axes of symmetry. A Patterson map calculated from the native X-ray data confirms that there are two dodecamers in the asymmetric unit, having both their sixfold and twofold axes parallel to one another.

scholarly journals Crystallization and preliminary X-ray crystallographic analysis of human myotubularin-related protein 1

2015 ◽  
Vol 71 (3) ◽  
pp. 261-265 ◽  
Author(s):  
Seoung Min Bong ◽  
Seung Won Yang ◽  
Ji-Woong Choi ◽  
Seung Jun Kim ◽  
Byung Il Lee
Keyword(s):  
Escherichia Coli ◽  
Polyethylene Glycol ◽  
Synchrotron Radiation ◽  
Crystallographic Analysis ◽  
Asymmetric Unit ◽  
Related Protein ◽  
Unit Cell Parameters ◽  
Solvent Content ◽  
X Ray ◽  
Cell Parameters

Myotubularin-related protein 1 is a phosphatase that dephosphorylates phospholipids such as phosphatidylinositol 3-phosphate or phosphatidylinositol 3,5-bisphosphate. In this study, human MTMR1 was overexpressed inEscherichia coli, purified and crystallized at 277 K using polyethylene glycol 20 000 as a precipitant. Diffraction data were collected to 2.0 Å resolution using synchrotron radiation. The crystals belonged to space groupP1, with unit-cell parametersa= 67.219,b= 96.587,c= 97.581 Å, α = 87.597, β = 86.072, γ = 77.327°. Assuming the presence of four molecules in the asymmetric unit, the calculated Matthews coefficient value was 2.61 Å3 Da−1and the corresponding solvent content was 52.9%.

Crystallization and preliminary X-ray analysis of the Escherichia coli UDP-MurNAc-tripeptide D-alanyl-D-alanine-adding enzyme (MurF)

1999 ◽  
Vol 55 (12) ◽  
pp. 2033-2034 ◽  
Author(s):  
Youwei Yan ◽  
Sanjeev Munshi ◽  
Ying Li ◽  
Kelly Ann D. Pryor ◽  
Frank Marsilio ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Asymmetric Unit ◽  
Hanging Drop ◽  
Data Set ◽  
Solvent Content ◽  
X Ray ◽  
Protein Mass ◽  
Cell Dimensions ◽  
Unit Cell Dimensions ◽  
Crystal Volume

Crystals of the Escherichia coli UDP-MurNAc-tripeptide D-Ala-D-Ala-adding protein (MurF), which catalyzes the formation of the last metabolite of the bacterial cell-wall building block, have been grown in hanging-drop vapor-diffusion trials using PEG 8K as a precipitating agent. The crystals belong to hexagonal space group P61 or P65, with unit-cell dimensions a = b = 74, c = 425 Å. The asymmetric unit contains two molecules, with a crystal volume per protein mass (Vm ) of 3.4 Å3 Da−1 and a solvent content of about 64% by volume. A native data set to 2.8 Å resolution has been obtained from a frozen crystal using a synchrotron X-ray source.

Metal-selenolate chemistry: stereochemistry of adamantane-type clusters of formula [(μ-SePh)6(MSePh)4]2− (M = Zn and Cd)

1992 ◽  
Vol 70 (3) ◽  
pp. 792-801 ◽  
Author(s):  
Jagadese J. Vittal ◽  
Philip A. W. Dean ◽  
Nicholas C. Payne
Keyword(s):  
Monoclinic Space Group ◽  
X Ray Diffraction ◽  
Bridging Ligands ◽  
Triclinic Space Group ◽  
Space Group ◽  
X Ray ◽  
Space Groups ◽  
X Ray Crystallography ◽  
Cell Dimensions ◽  
Minimum Number

The structures of three tetramethylammonium salts containing anions of formula [(μ-SePh)6(MSePh)4]2− (M = Zn and Cd) were determined by single crystal X-ray diffraction techniques. The Zn salt crystallizes in different space groups depending upon the solvent combination used in the synthesis. Thus crystals of (Me4N)2[Zn4(SePh)10], 1, grown from a mixture of methanol, acetonitrile, and acetone are triclinic, space group [Formula: see text] with cell dimensions a = 13.214(2), b = 23.859(2), c = 13.072(1) Å, α = 91.134(8), β = 113.350(8), γ = 79.865(9)°, and Z = 2. In the absence of acetone, a solvated crystal (Me4N)2[Zn4(SePh)10]•CH3CN, 2, is formed, which belongs to the monoclinic space group P21/n with a = 14.248(1), b = 39.722(2), c = 13.408(1) Å, β = 97.132(5)°, and Z = 4. The Cd salt (Me4N)2[Cd4(SePh)10], 3, crystallizes in the monoclinic space group P21/c, with a = 20.830(2), b = 14.282(1), c = 25.872(1) Å, β = 99.626(6)°, and Z = 4. These three salts are the first examples of homoleptic, tetranuclear selenolatometal(II) anions with (μ-Se)6M4 cages of adamantane-type stereochemistry. In each case the phenyl substituents of the bridging ligands adopt the configuration [aae, aae, aee, aee], which has the minimum number of two 1,3-axial–axial non-bonding substituent interactions. Keywords: selenolate complexes, synthesis, X-ray crystallography, isomerism, adamantane stereochemistry.

X-ray analysis of the haemoglobin molecule

1953 ◽  
Vol 141 (902) ◽  
pp. 67-69 ◽  
Keyword(s):  
Molecular Weight ◽  
Present Note ◽  
Bound Water ◽  
X Ray ◽  
Space Groups ◽  
Cell Dimensions ◽  
The Face ◽  
Simple Features ◽  
Haemoglobin Molecule ◽  
Water Contents

Met-, oxy- and carboxyhaemoglobin of horse crystallize with the space group C 2 , and two molecules in the unit cell which must therefore lie on twofold axes and must be identical in orientation. As has been shown by Perutz, the crystals can be obtained in at least six expanded or shrunk stages, with varying water contents. Furthermore, the water can be replaced by salt solution. The projection on the face is centrosymmetric, so that F(h0l) is real and a Fourier projection of the structure can be constructed if the signs of ( F ) can be determined. A comparison of the various shrinkage stages has made it possible to reduce the alternative ways of assigning the signs to a relatively small number, though it has not yet been found possible to obtain a unique solution. The present note reports the progress which has so far been made. The haemoglobins from different mammalian sources (horse, ox, sheep, man) appear to be very similar in their structure. Their similarity is shown by their having the same molecular weight and volume, by their optical properties, by their X-ray Patterson diagrams, and by the way they pack into the crystal structures. The cell dimensions and space groups of some dozen different crystalline types have been determined by Perutz and his collaborators. Four of these forms, including that mentioned above, have especially simple features of packing, and from this it is possible to deduce the overall dimension of the molecule. The protein mole­cule itself has a volume of 83000 Å 3 , a density of 0.43 electron per Å 3 , and approxi­mate dimensions 55 × 55 × 65 Å. It appears to be surrounded by a layer of bound water, into which salt does not penetrate, and which raises its volume to 115000 Å 3 .

Crystal data for two cis-o,o'-azodioxytoluene modifications

1982 ◽  
Vol 15 (2) ◽  
pp. 245-246 ◽  
Author(s):  
M. Azoulay ◽  
L. Trysberg
Keyword(s):  
Title Compound ◽  
Crystal Data ◽  
Crystal Forms ◽  
Orthorhombic System ◽  
Space Groups ◽  
Temperature Range ◽  
Compound C ◽  
Cell Dimensions

The title compound, C14H14N2O2, crystallizes from solutions in two different forms, α and β. The α form belongs to the orthorhombic system with cell dimensions a = 15.554(7), b = 15.451(8), c = 10.629(6) Å, V = 2.554(2) Å3 and Z = 8. Possible space groups are I m m a or I m 2 a. The β modification is monoclinic. P21/c, with a = 14.939(14), b = 9.587(5), c = 18.512(8) Å, β = 91.75(4)°, V = 2.598(3) Å3 and Z = 8. Indexed powder data are given for the two crystal forms. Within the temperature range investigated (255–298 K) the β form seems to be metastable.

Crystallization and preliminary X-ray studies of β-1,4-galactanase from Aspergillus aculeatus

1999 ◽  
Vol 55 (4) ◽  
pp. 929-930 ◽  
Author(s):  
C. Ryttersgaard ◽  
J.-C. N. Poulsen ◽  
S. Christgau ◽  
T. Sandal ◽  
H. Dalbøge ◽  
...  
Keyword(s):  
Asymmetric Unit ◽  
X Ray Diffraction ◽  
Aspergillus Aculeatus ◽  
Solvent Content ◽  
X Ray ◽  
Space Groups ◽  
Peg 400 ◽  
Cell Dimensions ◽  
Vapour Diffusion ◽  
Unit Cell Dimensions

Recombinant β-1,4-galactanase from Aspergillus aculeatus has been crystallized and characterized by X-ray diffraction. Crystals were obtained in hanging drops by vapour-diffusion under the conditions 30% PEG 400, 0.2 M CaCl2 and 0.1 M Na HEPES buffered to pH 7.5. The crystals diffract to 2.3 Å resolution and belong to one of the orthorhombic space groups I222 or I212121. The unit-cell dimensions are a = 60.42, b = 88.94 and c = 129.08 Å. With one molecule in the asymmetric unit, the corresponding solvent content is ∼48%.

scholarly journals Crystallization and preliminary X-ray diffraction analysis of the periplasmic domain of theEscherichia coliaspartate receptor Tar and its complex with aspartate

2014 ◽  
Vol 70 (9) ◽  
pp. 1219-1223 ◽  
Author(s):  
Takeshi Mise ◽  
Hideyuki Matsunami ◽  
Fadel A. Samatey ◽  
Ichiro N. Maruyama
Keyword(s):  
Escherichia Coli ◽  
Sodium Chloride ◽  
Molecular Mechanisms ◽  
Bacterial Chemotaxis ◽  
Cell Surface Receptor ◽  
X Ray Diffraction ◽  
X Ray ◽  
Space Groups ◽  
Surface Receptor ◽  
Periplasmic Domain

The cell-surface receptor Tar mediates bacterial chemotaxis toward an attractant, aspartate (Asp), and away from a repellent, Ni2+. To understand the molecular mechanisms underlying the induction of Tar activity by its ligands, theEscherichia coliTar periplasmic domain with and without bound aspartate (Asp-Tar and apo-Tar, respectively) were each crystallized in two different forms. Using ammonium sulfate as a precipitant, crystals of apo-Tar1 and Asp-Tar1 were grown and diffracted to resolutions of 2.10 and 2.40 Å, respectively. Alternatively, using sodium chloride as a precipitant, crystals of apo-Tar2 and Asp-Tar2 were grown and diffracted to resolutions of 1.95 and 1.58 Å, respectively. Crystals of apo-Tar1 and Asp-Tar1 adopted space groupP41212, while those of apo-Tar2 and Asp-Tar2 adopted space groupsP212121andC2, respectively.

Expression, purification, crystallization and preliminary X-ray analysis of Escherichia coli argininosuccinate synthetase

1999 ◽  
Vol 55 (12) ◽  
pp. 2028-2030 ◽  
Author(s):  
Chris Lemke ◽  
Melissa Yeung ◽  
P. Lynne Howell
Keyword(s):  
Escherichia Coli ◽  
National Laboratory ◽  
Brookhaven National Laboratory ◽  
Argininosuccinate Synthetase ◽  
Affinity Tag ◽  
X Ray ◽  
Diffusion Technique ◽  
Cell Dimensions ◽  
Synchrotron Light ◽  
Unit Cell Dimensions

A recombinant form of Escherichia coli argininosuccinate synthetase with a C-terminal polyhistidine affinity tag has been expressed, purified and subsequently crystallized using the hanging-drop vapour-diffusion technique. The crystals grow as large rectangular chunks with unit-cell dimensions a = 79.70, b = 105.84, c = 127.33 Å, α = β = γ = 90°. The crystals exhibit the symmetry of space group I222 and diffract to a minimum d-spacing of 1.6 Å at station X8C of the National Synchrotron Light Source, Brookhaven National Laboratory. On the basis of density calculations, one monomer of this homotetrameric protein is predicted per asymmetric unit (Matthews coefficient Vm = 2.69 Å3 Da−1).

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