scholarly journals Iodination of glyceraldehyde 3-phosphate dehydrogenase from Bacillus stearothermophilus

1976 ◽  
Vol 155 (3) ◽  
pp. 523-534 ◽  
Author(s):  
G Allen ◽  
J I Harris
Keyword(s):  
Conformational Changes ◽  
Bacillus Stearothermophilus ◽  
Three Dimensional ◽  
Thiol Group ◽  
Cysteine Residue ◽  
Enzymic Activity ◽  
Three Dimensional Model ◽  
Muscle Enzyme ◽  
Tyrosine Residues ◽  
Pig Muscle

The reaction of iodine with glyceraldehyde 3-phosphate dehydrogenase from Bacillus stearothermophilus was investigated. The active-site thiol group of the cysteine residue homologous with cysteine-149 in the pig muscle enzyme was protected by reaction with tetrathionate. The apoenzyme was readily inhibited by KI3 solution at pH8, but the coenzyme, NAD+, protected the enzyme against inhibition and decreased the extent of iodination. At pH 9.5, ready inhibition of both apo- and holo-enzyme was observed. Tryptic peptides containing residues iodinated at pH 8 were isolated and characterized. One of the most reactive residues in both holo- and apo-enzymes was a tyrosine homologous with tyrosine-46 in the pig muscle enzyme, and this residue was iodinated without loss of enzymic activity. Other reactive tyrosine residues in the apoenzyme were in positions homologous with residues 178, 273, 283 and 311 in the pig muscle enzyme, but they were not readily iodinated in the holoenzyme. Histidine residues in both holo- and apo-enzymes were iodinated at pH 8 in sequence positions homologous with residues 50, 162 and 190 in the pig muscle enzyme. The inhibition of the enzyme was not correlated with the iodination of a particular residue. The results are discussed in relation to a three-dimensional model based on the structure of the lobster muscle enzyme and demonstrate that conformational changes affecting the reactivity of several tyrosine residues most probably occur on binding of the coenzyme.

Characterization of the Mycobacterium tuberculosis H37Rv alkyl hydroperoxidase AhpC points to the importance of ionic interactions in oligomerization and activity

2001 ◽  
Vol 354 (1) ◽  
pp. 209-215 ◽  
Author(s):  
Radha CHAUHAN ◽  
Shekhar C. MANDE
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Conformational Changes ◽  
Three Dimensional ◽  
Enteric Bacteria ◽  
Enzymic Activity ◽  
Ionic Interactions ◽  
Three Dimensional Model ◽  
Mycobacterium Tuberculosis H37rv ◽  
Resistant Strains ◽  
Alkyl Hydroperoxidase

An alkyl hydroperoxidase (AhpC) has been found frequently to be overexpressed in isoniazid-resistant strains of Mycobacterium tuberculosis. These strains have an inactivated katG gene encoding a catalase peroxidase, which might render mycobacteria susceptible to the toxic peroxide radicals, thus leading to the concomitant overexpression of the AhpC. Although the overexpressed AhpC in isoniazid-resistant strains of M. tuberculosis may not directly participate in isoniazid action, AhpC might still assist M. tuberculosis in combating oxidative damage in the absence of the catalase. Here we have attempted to characterize the AhpC protein biochemically and report its functional and oligomerization properties. The alkyl hydroperoxidase of M. tuberculosis is unique in many ways compared with its well-characterized homologues from enteric bacteria. We show that AhpC is a decameric protein, composed of five identical dimers held together by ionic interactions. Dimerization of individual subunits takes place through an intersubunit disulphide linkage. The ionic interactions play a significant role in enzymic activity of the AhpC protein. The UV absorption spectrum and three-dimensional model of AhpC suggest that interesting conformational changes may take place during oxidation and reduction of the intersubunit disulphide linkage. In the absence of the partner AhpF subunit in M. tuberculosis, the mycobacterial AhpC might use small-molecule reagents, such as mycothiol, for completing its enzymic cycle.

scholarly journals Selectively Receptor-Blind Measles Viruses: Identification of Residues Necessary for SLAM- or CD46-Induced Fusion and Their Localization on a New Hemagglutinin Structural Model

2004 ◽  
Vol 78 (1) ◽  
pp. 302-313 ◽  
Author(s):  
Sompong Vongpunsawad ◽  
Numan Oezgun ◽  
Werner Braun ◽  
Roberto Cattaneo
Keyword(s):  
Amino Acids ◽  
Conformational Changes ◽  
Measles Virus ◽  
Structural Model ◽  
Three Dimensional ◽  
Lymphocyte Activation ◽  
Cell Receptor ◽  
Dimensional Model ◽  
Three Dimensional Model ◽  
H Protein

ABSTRACT Measles virus (MV) enters cells either through the signaling lymphocyte activation molecule SLAM (CD150) expressed only in immune cells or through the ubiquitously expressed regulator of complement activation, CD46. To identify residues on the attachment protein hemagglutinin (H) essential for fusion support through either receptor, we devised a strategy based on analysis of morbillivirus H-protein sequences, iterative cycles of mutant protein production followed by receptor-based functional assays, and a novel MV H three-dimensional model. This model uses the Newcastle disease virus hemagglutinin-neuraminidase protein structure as a template. We identified seven amino acids important for SLAM- and nine for CD46 (Vero cell receptor)-induced fusion. The MV H three-dimensional model suggests (i) that SLAM- and CD46-relevant residues are located in contiguous areas in propeller β-sheets 5 and 4, respectively; (ii) that two clusters of SLAM-relevant residues exist and that they are accessible for receptor contact; and (iii) that several CD46-relevant amino acids may be shielded from direct receptor contacts. It appears likely that certain residues support receptor-specific H-protein conformational changes. To verify the importance of the H residues identified with the cell-cell fusion assays for virus entry into cells, we transferred the relevant mutations into genomic MV cDNAs. Indeed, we were able to recover recombinant viruses, and we showed that these replicate selectively in cells expressing SLAM or CD46. Selectively receptor-blind viruses will be used to study MV pathogenesis and may have applications for the production of novel vaccines and therapeutics.

Study of the structure–activity relationships for the pyrazinamidase (PncA) from Mycobacterium tuberculosis

2001 ◽  
Vol 353 (3) ◽  
pp. 453-458 ◽  
Author(s):  
Nadine LEMAITRE ◽  
Isabelle CALLEBAUT ◽  
Frédéric FRENOIS ◽  
Vincent JARLIER ◽  
Wladimir SOUGAKOFF
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Three Dimensional ◽  
Cysteine Residue ◽  
Single Amino Acid ◽  
Three Dimensional Model ◽  
Mutant Proteins ◽  
Hydrophobic Cluster Analysis ◽  
Structural Modifications ◽  
Level Of Activity ◽  
Α Helix

In an attempt to investigate the molecular basis of pyrazinamide hydrolysis by the PncA protein from Mycobacterium tuberculosis, we determined the pyrazinamidase activity of nine PncA mutants bearing a single amino acid substitution. Among them, three mutants (D8G, K96T and S104R) had virtually no activity (⩽ 0.004unit/mg), five (F13S, T61P, P69L, Y103S and A146V) retained a low level of activity (0.06–0.25unit/mg) and one (T167L) exhibited a wild-type activity (1.51units/mg). The possible structural effects of these substitutions were assessed by analysing a three-dimensional model of the PncA protein constructed on the basis of the crystal structure of the N-carbamoylsarcosine amidohydrolase (CSHase) from Arthrobacter sp., an amidohydrolase which was found by hydrophobic cluster analysis to be closely related to PncA. In the PncA model, five of the mutated residues, Asp-8, Phe-13, Lys-96, Tyr-103 and Ser-104, were located within a 6 Å sphere around the cysteine residue Cys-138, which could be the counterpart of the active cysteine residue Cys-177 found in the CSHase. Among the remaining mutated residues, Thr-61, Pro-69 and Ala-146 were found to be more distant from Cys-138 but were associated with structural elements contributing to the catalytic centre, whereas Thr-167 was situated in an α-helix located far from the putative active site. These data suggest that the decrease in pyrazinamidase activity observed in the PncA mutant proteins is well correlated with the structural modifications the mutations can cause in the environment of the putative active cysteine Cys-138.

scholarly journals A study of the role of the reactive thiol group of rabbit muscle creatine kinase with a chromophoric reporter group

1978 ◽  
Vol 171 (1) ◽  
pp. 269-272 ◽  
Author(s):  
M A Keighren ◽  
N C Price
Keyword(s):  
Creatine Kinase ◽  
Conformational Changes ◽  
Thiol Group ◽  
Enzymic Activity ◽  
Rabbit Muscle ◽  
Muscle Creatine Kinase ◽  
Transition State Analogue ◽  
Muscle Creatine ◽  
Reporter Group

Substrate- and ligand-induced conformational changes were studied in a series of thiol-modified derivatives of rabbit muscle creatine kinase that retained different amounts of enzymic activity. The results indicate that the ‘reactive’ thiol group of the enzyme is required for the conformational changes associated with formation of a ‘transition-state analogue’ complex.

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).

Three-Dimensional Model for Virtual Surgical Simulation, during Complex Skull Base Surgery and Aneurysm Surgery

Skull Base ◽  
2008 ◽  
Vol 18 (S 01) ◽  
Author(s):  
Akio Morita ◽  
Toshikazu Kimura ◽  
Shigeo Sora ◽  
Kengo Nishimura ◽  
Hisayuki Sugiyama ◽  
...  
Keyword(s):  
Skull Base ◽  
Surgical Simulation ◽  
Skull Base Surgery ◽  
Three Dimensional ◽  
Aneurysm Surgery ◽  
Dimensional Model ◽  
Three Dimensional Model

Digitalization system of ancient architecture decoration art based on neural network and image features

2020 ◽  
pp. 1-12
Author(s):  
Wu Xin ◽  
Qiu Daping
Keyword(s):  
Neural Network ◽  
Construction Industry ◽  
Three Dimensional ◽  
Performance Testing ◽  
Image Features ◽  
Three Dimensional Model ◽  
Performance Effect ◽  
Data Process ◽  
And Performance ◽  
Construction Mode

The inheritance and innovation of ancient architecture decoration art is an important way for the development of the construction industry. The data process of traditional ancient architecture decoration art is relatively backward, which leads to the obvious distortion of the digitalization of ancient architecture decoration art. In order to improve the digital effect of ancient architecture decoration art, based on neural network, this paper combines the image features to construct a neural network-based ancient architecture decoration art data system model, and graphically expresses the static construction mode and dynamic construction process of the architecture group. Based on this, three-dimensional model reconstruction and scene simulation experiments of architecture groups are realized. In order to verify the performance effect of the system proposed in this paper, it is verified through simulation and performance testing, and data visualization is performed through statistical methods. The result of the study shows that the digitalization effect of the ancient architecture decoration art proposed in this paper is good.

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