Mapping of monoclonal antibodies specific to P64k: A common antigen of several isolates of Neisseria meningitidis

2001 ◽  
Vol 47 (2) ◽  
pp. 158-164 ◽  
Author(s):  
C Nazábal ◽  
T Carmenate ◽  
S Cruz ◽  
S González ◽  
R Silva ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Neisseria Meningitidis ◽  
Three Dimensional ◽  
Structural Data ◽  
Dimensional Structure ◽  
Bacterial Surface ◽  
A Minor ◽  
The Relationship ◽  
Cell Elisa ◽  
Competition Assays

P64k is a minor outer membrane protein from Neisseria meningitidis. This protein has been produced at high levels in Escherichia coli. We generated a group of monoclonal antibodies (mAbs) against recombinant P64k, which recognise four non-overlapping epitopes, as shown using competition assays with biotinylated mAbs. The P64k sequences involved in mAbs binding were mapped with synthetic overlapping peptides derived from the P64k protein, and located in the previously determined three-dimensional structure of the protein. These antibodies were also characterised by whole-cell ELISA and bactericidal tests against N. meningitidis. Only two of the recognised epitopes were exposed on the bacterial surface, and none of the mAbs showed bactericidal activity. The relationship between these results and the structural data on the epitopes bound by the mAbs is discussed.Key words: Neisseria meningitidis, P64k, monoclonal antibodies, epitope mapping.

Characterization of Monoclonal Anti-human Factor VII Antibody (B101/B1) that Recognized Three-dimensional Structures near Arg at Position 79 in the First EGF-like Domain

1998 ◽  
Vol 79 (01) ◽  
pp. 104-109 ◽  
Author(s):  
Osamu Takamiya
Keyword(s):  
Monoclonal Antibodies ◽  
Tissue Factor ◽  
Human Factor ◽  
Solid Phase ◽  
Three Dimensional ◽  
Coagulation Factors ◽  
Factor Vii ◽  
Dimensional Structure ◽  
Epidermal Growth

SummaryMurine monoclonal antibodies (designated hVII-B101/B1, hVIIDC2/D4 and hVII-DC6/3D8) directed against human factor VII (FVII) were prepared and characterized, with more extensive characterization of hVII-B101/B1 that did not bind reduced FVIIa. The immunoglobulin of the three monoclonal antibodies consisted of IgG1. These antibodies did not inhibit procoagulant activities of other vitamin K-dependent coagulation factors except FVII and did not cross-react with proteins in the immunoblotting test. hVII-DC2/D4 recognized the light chain after reduction of FVIIa with 2-mercaptoethanol, and hVIIDC6/3D8 the heavy chain. hVII-B101/B1 bound FVII without Ca2+, and possessed stronger affinity for FVII in the presence of Ca2+. The Kd for hVII-B101/B1 to FVII was 1.75 x 10–10 M in the presence of 5 mM CaCl2. The antibody inhibited the binding of FVII to tissue factor in the presence of Ca2+. hVII-B101/B1 also inhibited the activation of FX by the complex of FVIIa and tissue factor in the presence of Ca2+. Furthermore, immunoblotting revealed that hVII-B101/B1 reacted with non-reduced γ-carboxyglutaminic acid (Gla)-domainless-FVII and/or FVIIa. hVII-B101/B1 showed a similar pattern to that of non-reduced proteolytic fragments of FVII by trypsin with hVII-DC2/D4 on immunoblotting test. hVII-B101/B1 reacted differently with the FVII from the dysfunctional FVII variant, FVII Shinjo, which has a substitution of Gln for Arg at residue 79 in the first epidermal growth factor (1st EGF)-like domain (Takamiya O, et al. Haemosta 25, 89-97,1995) compared with normal FVII, when used as a solid phase-antibody for ELISA by the sandwich method. hVII-B101/B1 did not react with a series of short peptide sequences near position 79 in the first EGF-like domain on the solid-phase support for epitope scanning. These results suggested that the specific epitope of the antibody, hVII-B101/B1, was located in the three-dimensional structure near position 79 in the first EGF-like domain of human FVII.

Determining the orientation of parent β grain from one α variant in titanium alloys

2018 ◽  
Vol 51 (4) ◽  
pp. 1125-1132 ◽  
Author(s):  
Z. B. Zhao ◽  
Q. J. Wang ◽  
H. Wang ◽  
J. R. Liu ◽  
R. Yang
Keyword(s):  
Titanium Alloys ◽  
Habit Plane ◽  
Electron Backscatter Diffraction ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Plane Normal ◽  
Β Phase ◽  
Microstructural Morphology ◽  
Backscatter Diffraction ◽  
The Relationship

The relationship between the crystallographic orientation and habit plane normal of transformed α laths in titanium alloys is discussed according to the Burgers orientation relationship and the three-dimensional structure of the α lath. A new method (orientation–trace method) is developed to determine the orientation of the parent β phase using the orientation of the α lath, which was measured by electron backscatter diffraction, and the microstructural morphology of that α variant. This approach is validated in a near-α titanium alloy. Moreover, the habit plane normal direction of the transformed α lath can be obtained from the crystallographic orientations of the α lath itself and its parent β grain. The verification and the corresponding discussion show the reliability of this approach.

scholarly journals SphereCon—a method for precise estimation of residue relative solvent accessible area from limited structural information

2020 ◽  
Vol 36 (11) ◽  
pp. 3372-3378
Author(s):  
Alexander Gress ◽  
Olga V Kalinina
Keyword(s):  
Protein Function ◽  
Structural Information ◽  
Solvent Accessibility ◽  
Three Dimensional ◽  
Structural Data ◽  
Supplementary Information ◽  
Dimensional Structure ◽  
Relative Solvent Accessibility ◽  
Precise Measure ◽  
The Impact

Abstract Motivation In proteins, solvent accessibility of individual residues is a factor contributing to their importance for protein function and stability. Hence one might wish to calculate solvent accessibility in order to predict the impact of mutations, their pathogenicity and for other biomedical applications. A direct computation of solvent accessibility is only possible if all atoms of a protein three-dimensional structure are reliably resolved. Results We present SphereCon, a new precise measure that can estimate residue relative solvent accessibility (RSA) from limited data. The measure is based on calculating the volume of intersection of a sphere with a cone cut out in the direction opposite of the residue with surrounding atoms. We propose a method for estimating the position and volume of residue atoms in cases when they are not known from the structure, or when the structural data are unreliable or missing. We show that in cases of reliable input structures, SphereCon correlates almost perfectly with the directly computed RSA, and outperforms other previously suggested indirect methods. Moreover, SphereCon is the only measure that yields accurate results when the identities of amino acids are unknown. A significant novel feature of SphereCon is that it can estimate RSA from inter-residue distance and contact matrices, without any information about the actual atom coordinates. Availability and implementation https://github.com/kalininalab/spherecon. Contact [email protected] Supplementary information Supplementary data are available at Bioinformatics online.

The magnetotelluric impedance tensor properties with respect to rotations

Geophysics ◽  
1985 ◽  
Vol 50 (10) ◽  
pp. 1610-1617 ◽  
Author(s):  
Simon Spitz
Keyword(s):  
Data Analysis ◽  
Coordinate System ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Impedance Tensor ◽  
Coordinate Systems ◽  
Magnetotelluric Data ◽  
Conventional Analysis ◽  
Complete Set ◽  
The Relationship

A serious limitation to conventional data analysis is that the data refer mainly to elongated bodies. When three‐dimensional distortions are present, quantitative interpretation based only on the off‐diagonal elements of the conventionally rotated impedance tensor is inadequate, because these off‐diagonal elements are insensitive to the tensor trace. The impedance tensor eigenstate formulation proposed in the literature defines a complete set of parameters suitable for recognition of three‐dimensionality. Generally, though, the eigenvalues do not stand for the off‐diagonal elements of an impedance tensor measured in a physical coordinate system. It is shown how the eigenvalues are modified when the relationship between coordinate system rotations and the eigenstate formulation is clarified. A generalization of the conventional analysis results, but the rotation angle obtained is neither unique nor complete To improve the situation, two new analytical rotation angles are proposed. These angles define two complete intrinsic coordinate systems suitable for magnetotelluric data analysis when a general three‐dimensional structure is involved.

scholarly journals Crystal structure ofN-(quinolin-6-yl)hydroxylamine

2014 ◽  
Vol 70 (11) ◽  
pp. 322-324 ◽  
Author(s):  
Anuruddha Rajapakse ◽  
Roman Hillebrand ◽  
Sarah M. Lewis ◽  
Zachary D. Parsons ◽  
Charles L. Barnes ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Hydrogen Bonds ◽  
Title Compound ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Asymmetric Unit ◽  
Π Interactions ◽  
Compound C ◽  
Independent Molecules ◽  
A Minor

The title compound, C9H8N2O, crystallized with four independent molecules in the asymmetric unit. The four molecules are linkedviaone O—H...N and two N—H...N hydrogen bonds, forming a tetramer-like unit. In the crystal, molecules are further linked by O—H...N and N—H...O hydrogen bonds forming layers parallel to (001). These layers are linkedviaC—H...O hydrogen bonds and a number of weak C—H...π interactions, forming a three-dimensional structure. The crystal was refined as a non-merohedral twin with a minor twin component of 0.319.

scholarly journals Three-Dimensional Structure of the Neisseria meningitidis Secretin PilQ Determined from Negative-Stain Transmission Electron Microscopy

2003 ◽  
Vol 185 (8) ◽  
pp. 2611-2617 ◽  
Author(s):  
Richard F. Collins ◽  
Robert C. Ford ◽  
Ashraf Kitmitto ◽  
Ranveig O. Olsen ◽  
Tone Tønjum ◽  
...  
Keyword(s):  
Neisseria Meningitidis ◽  
Outer Membrane ◽  
Protein Complex ◽  
Rotational Symmetry ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Three Dimensional Structure ◽  
Type Iv ◽  
Negative Stain ◽  
Dominant Feature

ABSTRACT The PilQ secretin from the pathogenic bacterium Neisseria meningitidis is an integral outer membrane protein complex which plays a crucial role in the biogenesis of type IV pili. We present here the first three-dimensional structure of this type of secretin at 2.5-nm resolution, obtained by single-particle averaging methods applied to the purified protein complex visualized in a negative stain. In projection, the PilQ complex is circular, with a donut-like appearance. When viewed from the side it has a rounded, conical profile. The complex was demonstrated to have 12-fold rotational symmetry, and this property was used to improve the quality of the density map by symmetry averaging. The dominant feature of the structure is a cavity, 10 nm deep, within the center of the molecule. The cavity is funnel-shaped in cross section, measures 6.5 nm in diameter at the top of the complex, and tapers to a closed point, effectively blocking formation of a continuous pore through the PilQ complex. These results suggest that the complex would have to undergo a conformational change in order to accommodate an assembled pilus fiber of diameter 6.5 nm running through the outer membrane.

scholarly journals Older workers: stereotypes and occupational self-efficacy

2016 ◽  
Vol 31 (7) ◽  
pp. 1152-1166 ◽  
Author(s):  
Rita Chiesa ◽  
Stefano Toderi ◽  
Paola Dordoni ◽  
Kene Henkens ◽  
Elena Maria Fiabane ◽  
...  
Keyword(s):  
Older Workers ◽  
Self Efficacy ◽  
Age Groups ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Age Stereotypes ◽  
Content Type ◽  
Moderator Role ◽  
The Relationship ◽  
Organizational Age

Purpose The purpose of this paper is to explore the relationship between organizational age stereotypes and occupational self-efficacy. First, the authors intend to test the measurement invariance of Henkens’s (2005) age stereotypes scale across two age group, respectively, under 50 and 50 years and older. Then, the moderator role of age groups in the relationship between age stereotypes and occupational self-efficacy is investigated. Design/methodology/approach The survey involved a large sample of 4,667 Italian bank sector’s employees. Findings The results show the invariance of the three dimensional structure of organizational stereotypes towards older workers scale: productivity, reliability and adaptability. Furthermore, the moderation is confirmed: the relationship between organizational age stereotypes and occupational self-efficacy is significant only for older respondents. Research limitations/implications Future studies should aim to replicate the findings with longitudinal designs. Practical implications The study suggests the importance to emphasize the positive characteristics of older workers and to reduce the presence of negative age stereotypes in the workplace, especially in order to foster the occupational self-efficacy of older workers. Originality/value The findings are especially relevant in view of the lack of evidence about the relationship between age stereotypes and occupational self-efficacy.

Three-dimensional structure of a regular bacterial surface layer: The HPI-layer of Deinococcus radiodurans

1983 ◽  
Vol 41 ◽  
pp. 438-439
Author(s):  
W. Baumeister ◽  
M. Hahn ◽  
W.O. Saxton
Keyword(s):  
Outer Membrane ◽  
Protein Interactions ◽  
Deinococcus Radiodurans ◽  
Bacterial Species ◽  
Hexagonal Lattice ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Protein Protein Interactions ◽  
Bacterial Surface ◽  
Freeze Dried

Regularly organized surface (RS) layers are a feature common to many bacterial species; they are clearly more abundant than was anticipated even a few years ago. The RS-layers are believed to fulfil a variety of functions in the interaction between the cell and its environment (see e.g. [1]). The so-called HPI-layer of the radiotolerant bacterium Deinococcus radiodurans is a typical example of such a layer: It is composed of a single polypeptide species (Mr 105 kDa) arranged on a hexagonal lattice to form a network that covers the entire surface of the bacterium; it is associated with the outer membrane via hydrophobic protein-protein interactions.Isolated HPI-layer sheets, released from the outer membrane by detergent treatment, have been studied in the electron microscope making extensive use of the present arsenal of preparation techniques: negative staining, (auro- thio)glucose embedding, freeze-dried/unstained, freeze-dried/metal shadowed etc.Because of the notorious problem of lattice imperfections image processing usually followed the strategy of correlation averaging as outlined in some detail elsewhere.

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