scholarly journals Quaternary structure ofDioclea grandifloralectin assessed by equilibrium sedimentation and crystallographic analysis of recombinant mutants

FEBS Letters ◽  
2015 ◽  
Vol 589 (18) ◽  
pp. 2290-2296 ◽  
Author(s):  
Sara Zamora-Caballero ◽  
Alicia Pérez ◽  
Libia Sanz ◽  
Jerónimo Bravo ◽  
Juan J. Calvete
Keyword(s):  
Quaternary Structure ◽  
Crystallographic Analysis ◽  
Equilibrium Sedimentation

Quaternary structure of Limulus polyphemus hemocyanin

1978 ◽  
Vol 36 (2) ◽  
pp. 176-177
Author(s):  
T. Wichertjes ◽  
E.J. Kwak ◽  
E.F.J. Van Bruggen
Keyword(s):  
Electron Microscopy ◽  
Functional Properties ◽  
Horseshoe Crab ◽  
Temperature Jump ◽  
Quaternary Structure ◽  
Crystallographic Analysis ◽  
Limulus Polyphemus ◽  
Oxygen Binding ◽  
X Ray ◽  
Intact Molecule

Hemocyanin of the horseshoe crab (Limulus polyphemus) has been studied in nany ways. Recently the structure, dissociation and reassembly was studied using electron microscopy of negatively stained specimens as the method of investigation. Crystallization of the protein proved to be possible and X-ray crystallographic analysis was started. Also fluorescence properties of the hemocyanin after dialysis against Tris-glycine buffer + 0.01 M EDTA pH 8.9 (so called “stripped” hemocyanin) and its fractions II and V were studied, as well as functional properties of the fractions by NMR. Finally the temperature-jump method was used for assaying the oxygen binding of the dissociating molecule and of preparations of isolated subunits. Nevertheless very little is known about the structure of the intact molecule. Schutter et al. suggested that the molecule possibly consists of two halves, combined in a staggered way, the halves themselves consisting of four subunits arranged in a square.

Crystallization and preliminary crystallographic analysis of the major horse allergen Equ c 1

1999 ◽  
Vol 55 (4) ◽  
pp. 880-882 ◽  
Author(s):  
Christophe Grégoire ◽  
Gisele A. Tavares ◽  
Hans K. Lorenzo ◽  
Jean-Pierre Dandeu ◽  
Bernard David ◽  
...  
Keyword(s):  
Single Molecule ◽  
Gel Filtration ◽  
Specific Ige ◽  
Crystallographic Analysis ◽  
Recombinant Form ◽  
Data Set ◽  
Synchrotron Source ◽  
Cell Parameters ◽  
Low Protein ◽  
Equilibrium Sedimentation

The secreted protein Equ c 1 is the major component responsible for the induction of specific IgE antibodies in patients sensitized to horse allergens. Equ c 1 belongs to the lipocalin superfamily of hydrophobic ligand-binding proteins, which also includes other known allergens. Equilibrium sedimentation and gel-filtration studies demonstrate that both the glycosylated form of Equ c 1 purified from horse salivary glands and the non-glycosylated recombinant form expressed in bacteria exist predominantly as dimers in solution. As observed for other dimeric lipocalins, acidic pH and low protein concentration favour dimer dissociation. The recombinant form of Equ c 1 has been crystallized using ammonium sulfate as a precipitant. The crystals belong to the tetragonal space group P41212 with cell parameters a = b = 84.0, c = 56.1 Å, and contain a single molecule in the asymmetric unit. A complete data set from native crystals was collected at the synchrotron source in Hamburg to 2.9 Å resolution using a frozen crystal, and structure determination is in progress.

Crystallization and preliminary crystallographic analysis of the hydroxyquinol 1,2-dioxygenase from Nocardioides simplex 3E: a novel dioxygenase involved in the biodegradation of polychlorinated aromatic compounds

1999 ◽  
Vol 55 (4) ◽  
pp. 901-903 ◽  
Author(s):  
Manuela Benvenuti ◽  
Fabrizio Briganti ◽  
Andrea Scozzafava ◽  
Ludmilla Golovleva ◽  
Vasily M. Travkin ◽  
...  
Keyword(s):  
Quaternary Structure ◽  
Crystallographic Analysis ◽  
Aerobic Biodegradation ◽  
Orthorhombic Space Group ◽  
Data Set ◽  
Cell Parameters ◽  
Peg 400 ◽  
Laboratory Source ◽  
Intradiol Cleavage ◽  
Intradiol Dioxygenase

Hydroxyquinol 1,2-dioxygenase (HQ1,2O) from Nocardioides simplex 3E, an enzyme involved in the aerobic biodegradation of a large class of chloroaromatic compounds such as 2,4-dichlorophenoxyacetate (2,4-D) and 2,4,5-trichlorophenoxyacetate (2,4,5-T), has been crystallized. HQ1,2O, which specifically catalyzes the intradiol cleavage of hydroxyquinol (1,2,4-trihydroxybenzene), an intermediate in the degradation of a variety of aromatic pollutants, to maleylacetate, has been recently purified to homogeneity. The enzyme is an homodimer composed of two identical subunits in a α2-type quaternary structure, has a molecular weight of about 65 kDa and contains a catalytically essential Fe(III) ion. Crystals of HQ1,2O obtained using 2% PEG 400 and 2 M ammonium sulfate at pH 7.5 as precipitants belong to the orthorhombic space group P212121, with unit-cell parameters a = 81.15 (6), b = 86.79 (7), c = 114.93 (8). Assuming one dimer per asymmetric unit, the Vm value is 2.51 Å3 Da−1. A complete native data set to 1.8 Å resolution has been collected on a laboratory source. This is the first intradiol dioxygenase which specifically catalyzes the cleavage of hydroxyquinol to give diffraction-quality crystals.

scholarly journals Insights into the structural basis of the pH-dependent dimer–tetramer equilibrium through crystallographic analysis of recombinant Diocleinae lectins

2007 ◽  
Vol 409 (2) ◽  
pp. 417-428 ◽  
Author(s):  
Celso S. Nagano ◽  
Juan J. Calvete ◽  
Domingo Barettino ◽  
Alicia Pérez ◽  
Benildo S. Cavada ◽  
...  
Keyword(s):  
Structural Information ◽  
Binding Activity ◽  
Crystallographic Analysis ◽  
Structural Basis ◽  
Carbohydrate Binding ◽  
Central Cavity ◽  
Legume Lectin ◽  
Equilibrium Sedimentation ◽  
Ph Dependency ◽  
Ph Dependent

The structural ground underlying the pH-dependency of the dimer–tetramer transition of Diocleinae lectins was investigated by equilibrium sedimentation and X-ray crystal structure determination of wild-type and site-directed mutants of recombinant lectins. Synthetic genes coding for the full-length α-chains of the seed lectins of Dioclea guianensis (termed r-αDguia) and Dioclea grandiflora (termed r-αDGL) were designed and expressed in Escherichia coli. This pioneering approach, which will be described in detail in the present paper, yielded recombinant lectins displaying carbohydrate-binding activity, dimer–tetramer equilibria and crystal structures indistinguishable from their natural homologues. Conversion of the pH-stable tetrameric r-αDGL into a structure exhibiting pH-dependent dimer–tetramer transition was accomplished through mutations that abolished the interdimeric interactions at the central cavity of the tetrameric lectins. Both the central and the peripheral interacting regions bear structural information for formation of the canonical legume lectin tetramer. We hypothesize that the strength of the ionic contacts at these sites may be modulated by the pH, leading to dissociation of those lectin structures that are not locked into a pH-stable tetramer through interdimeric contacts networking the central cavity loops.

Some Observations on Intra-Mitochondrial Crystals

1977 ◽  
Vol 35 ◽  
pp. 406-407
Author(s):  
J. A. Clarke ◽  
D. N. Landon ◽  
P. R. Ward
Keyword(s):  
Cytochrome B ◽  
Mitochondrial Myopathy ◽  
Crystallographic Analysis ◽  
Mitochondrial Membranes ◽  
Electron Microscopes ◽  
Muscle Biopsies

Intra-mitochondrial crystals have been noted in muscle biopsies from patients in a wide variety of diseased states. As far as we are aware, none of these crystals have been subjected to detailed crystallographic analysis. Recently, similar crystals were observed in a biopsy from a patient with a mitochondrial myopathy, characterised by a deficiency in reducible cytochrome b (Morgan-Hughes, J. A., Darveniza, P., Kahn, S. N., Landon, D. N., Sherratt, R. M., Land, J. M. and Clark, J. B., 1977, Brain, In Press). Aldehyde-fixed, osmicated resin imbedded material was examined using Siemens, JEOL and Phillips electron microscopes with goniometer specimen stages. The crystals generally lay between the outer and inner mitochondrial membranes and measured 1 - 3 μm in length and 0.1 - 0.3 μm in width. Characteristically, these crystals revealed specific periodicities.

Characterization of Tamm-Horsfall Urinary Glycoprotein

1973 ◽  
Vol 31 ◽  
pp. 420-421
Author(s):  
John P. Robinson ◽  
J. David Puett
Keyword(s):  
Electron Microscopy ◽  
Circular Dichroism ◽  
Secondary Structure ◽  
Quaternary Structure ◽  
Normal Adult ◽  
Morphological Properties ◽  
Adult Males ◽  
Circular Dichroïsm ◽  
Urinary Glycoprotein

Much work has been reported on the chemical, physical and morphological properties of urinary Tamm-Horsfall glycoprotein (THG). Although it was once reported that cystic fibrotic (CF) individuals had a defective THG, more recent data indicate that THG and CF-THG are similar if not identical.No studies on the conformational aspects have been reported on this glycoprotein using circular dichroism (CD). We examined the secondary structure of THG and derivatives under various conditions and have correlated these results with quaternary structure using electron microscopy.THG was prepared from normal adult males and CF-THG from a 16-year old CF female by the method of Tamm and Horsfall. CF female by the method of Tamm and Horsfall.

Electron Microscopy and image reconstruction reveal the structural basis for spectrin's elastic properties

1992 ◽  
Vol 50 (1) ◽  
pp. 510-511
Author(s):  
Amy M. McGough ◽  
Robert Josephs
Keyword(s):  
Electron Microscopy ◽  
Elastic Properties ◽  
Conformational Changes ◽  
Quaternary Structure ◽  
Three Dimensional ◽  
Membrane Skeleton ◽  
Projection Algorithm ◽  
Structural Basis ◽  
Iterative Approximation ◽  
Membrane Skeletons

The remarkable deformability of the erythrocyte derives in large part from the elastic properties of spectrin, the major component of the membrane skeleton. It is generally accepted that spectrin's elasticity arises from marked conformational changes which include variations in its overall length (1). In this work the structure of spectrin in partially expanded membrane skeletons was studied by electron microscopy to determine the molecular basis for spectrin's elastic properties. Spectrin molecules were analysed with respect to three features: length, conformation, and quaternary structure. The results of these studies lead to a model of how spectrin mediates the elastic deformation of the erythrocyte.Membrane skeletons were isolated from erythrocyte membrane ghosts, negatively stained, and examined by transmission electron microscopy (2). Particle lengths and end-to-end distances were measured from enlarged prints using the computer program MACMEASURE. Spectrin conformation (straightness) was assessed by calculating the particles’ correlation length by iterative approximation (3). Digitised spectrin images were correlation averaged or Fourier filtered to improve their signal-to-noise ratios. Three-dimensional reconstructions were performed using a suite of programs which were based on the filtered back-projection algorithm and executed on a cluster of Microvax 3200 workstations (4).

Morphology of Σ3{/70.53°} grain boundaries in a C15 intermetallic compound

1994 ◽  
Vol 52 ◽  
pp. 684-685
Author(s):  
Fuming Chu ◽  
D. P. Pope ◽  
D. S. Zhou ◽  
T. E. Mitchell
Keyword(s):  
Grain Boundary ◽  
Grain Boundaries ◽  
Crystallographic Analysis ◽  
Laves Phase ◽  
Second Phase ◽  
Bright Field Image ◽  
Boundary Edge ◽  
Arc Melting ◽  
Fcc Lattice ◽  
Diffraction Patterns

A C15 Laves phase, HfV2+Nb, shows promising mechanical properties and here we describe the structure of its grain boundaries. The C15 Laves phase has a fcc lattice with a=7.4Å. An alloy of composition Hf14V64Nb22 (including a C15 matrix and a second phase of V-rich bcc solution) was made by arc-melting. The alloy was homogenized at 1200°C for 120h. Preliminary study concentrated on Σ3{<110>/70.53°} grain boundaries in the C15 phase using Philips 400T and CM 30 microscopes.The most-commonly observed morphology of Σ3{<110>/70.53°} grain boundaries in the C15 phase is a faceted boundary. A bright field image (BFI) of the faceted boundary and the corresponding diffraction patterns with the grain boundary edge-on are shown in Fig. 1(a). From the diffraction patterns using a <110> zone axis for both grains, it is obvious that this is a Σ3{<110>/70.53°} grain boundary. Crystallographic analysis shows that the Σ3{<110>/70.53°} grain boundaries selectively facet with the following relationships between the two grains: {111}1//{111}2, {112}1//{112}2, {111}1//{115}2, and {001}1//{221}2.

A crystallographic analysis of the CoSi/Si(111) interface using Transmission Electron Microscopy

1990 ◽  
Vol 48 (4) ◽  
pp. 574-575
Author(s):  
A.C. Daykin ◽  
C.J. Kiely ◽  
R.C. Pond ◽  
J.L. Batstone
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Defect Structure ◽  
Room Temperature ◽  
Theoretical Method ◽  
Crystallographic Analysis ◽  
Si Substrate ◽  
Transmission Electron ◽  
Theoretical Predictions

When CoSi2 is grown onto a Si(111) surface it can form in two distinct orientations. A-type CoSi2 has the same orientation as the Si substrate and B-type is rotated by 180° degrees about the [111] surface normal.One method of producing epitaxial CoSi2 is to deposit Co at room temperature and anneal to 650°C.If greater than 10Å of Co is deposited then both A and B-type CoSi2 form via a number of intermediate silicides .The literature suggests that the co-existence of A and B-type CoSi2 is in some way linked to these intermediate silicides analogous to the NiSi2/Si(111) system. The phase which forms prior to complete CoSi2 formation is CoSi. This paper is a crystallographic analysis of the CoSi2/Si(l11) bicrystal using a theoretical method developed by Pond. Transmission electron microscopy (TEM) has been used to verify the theoretical predictions and to characterise the defect structure at the interface.

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