Insights into the Regulatory Characteristics of the Mycobacterial Dephosphocoenzyme A Kinase: Implications for the Universal CoA Biosynthesis Pathway

Being vastly different from the human counterpart, we suggest that the last enzyme of the Mycobacterium tuberculosis Coenzyme A biosynthetic pathway, dephosphocoenzyme A kinase (CoaE) could be a good anti-tubercular target. Here we describe detailed investigations into the regulatory features of the enzyme, affected via two mechanisms. Enzymatic activity is regulated by CTP which strongly binds the enzyme at a site overlapping that of the leading substrate, dephosphocoenzyme A (DCoA), thereby obscuring the binding site and limiting catalysis. The organism has evolved a second layer of regulation by employing a dynamic equilibrium between the trimeric and monomeric forms of CoaE as a means of regulating the effective concentration of active enzyme. We show that the monomer is the active form of the enzyme and the interplay between the regulator, CTP and the substrate, DCoA, affects enzymatic activity. Detailed kinetic data have been corroborated by size exclusion chromatography, dynamic light scattering, glutaraldehyde crosslinking, limited proteolysis and fluorescence investigations on the enzyme all of which corroborate the effects of the ligands on the enzyme oligomeric status and activity. Cysteine mutagenesis and the effects of reducing agents on mycobacterial CoaE oligomerization further validate that the latter is not cysteine-mediated or reduction-sensitive. These studies thus shed light on the novel regulatory features employed to regulate metabolite flow through the last step of a critical biosynthetic pathway by keeping the latter catalytically dormant till the need arises, the transition to the active form affected by a delicate crosstalk between an essential cellular metabolite (CTP) and the precursor to the pathway end-product (DCoA).

Beta-structure of polypeptides in non-aqueous solutions. I. Spectral characteristics of the polypeptide backbone

The optical rotatory features of the beta-structure of the polypeptides in non-aqueous solutions and films cast from these solutions have been investigated. The beta-structure of poly-S-benzyl-L-cysteine, poly-S-carbobenzoxy-L-cysteine and poly-S-benzyl-L-cysteine, poly-S-carbobenzoxy-L-cysteine and poly-O-carbo-bands of their films. The optical rotatory dispersion (ORD) and circular dichroism (CD) spectra of these polypeptides are found to be very similar in both film and solution. In solvents promoting the beta-structure, the polypeptides are characterized by CD troughs in the n-pi* transition region of the peptide chromophore. The ORD spectra are found to be positive in sign throughout the visible and accessible ultraviolet regions and are interpreted in terms of the possible existence of a relatively much larger positive pi-pi* CD bands as compared with the negative n-pi* band. The rotatory data obtained in the non-aqueous solution are compared with those obtained for other poly peptides in aqueous solutions, with respect to the type and extent of beta-structure present.

Influence of the Processing Conditions on the Characteristics of the Clad Layers Produced with Laminar Plasma Technology

Laminar plasma technology was used to produce ceramic hardened layers of Al2O3-40% mass Ni composite powders on stainless steel substrates. In order to investigate the influences of processing conditions on the morphologies of the surface modified layers, two different powder-feeding methods were tested, one with carrier gas called the powder injection method, and the other without carrier gas called powder transfers method. The microscopic investigations demonstrate that the cross-section of the clad layers consists of two distinct microstructural regions, in which the Al2O3 phases exhibit different growth mechanisms. When the powder transfers method is adopted, the number density and volume fraction of the Al2O3 particles increase considerably and their distributions exhibit zonal periodical characteristics. When the powder-feeding rate increases, the microstructure of the Al2O3 phases changes from a small globular to a long needle shape. Finite element simulations show that the transient thermo-physical features of the pool substances, such as solidification rate and cooling rate, influence strongly the mechanisms of the nucleation and the directional growth of the Al2O3 phases in the thermal processing.

Characteristics of the interface of a laser-quenched steel substrate and chromium electroplate

The interface of a laser-discrete-quenched steel substrate and as-deposited chromium electroplate was investigated by ion beam etching, dissolving-substrate-away and using a Vickers microhardness tester, in an attempt to reveal the mechanism that the service life of the chromium-coated parts is increased by the duplex technique of laser pre-quenching plus chromium post-depositing. The laser quenching of the steel substrate can reduce the steep hardness gradient at the substrate/chromium interface and improve the load-bearing capacity of chromium electroplate. Moreover, the laser quenching prior to plating has an extremely great effect on the morphologies and microstructure of the substrate/chromium interface: there is a transient interlayer at the original substrate/chromium interface while there is not at the laser-quenchedzone/chromium interface; the near-substrate surface microstructure and morphologies of the free-standing chromium electrodeposits, whose substrate was dissolved away with nital 30% in volume, inherit the periodically gradient characteristics of the laser-discrete-quenched substrate surface. (c) 2006 Elsevier B.V. All rights reserved.

Flow and chemical characteristics of the St. Johns River at Jacksonville, Florida

This report will be of substantial value to water managers in developing the St. Johns River as a multiple resource. Evaluation of the capacity of the river to accept pollutants without adversely affecting other uses requires detailed data of flow and chemical characteristics and an understanding of how they interact. (66 page document)

Investigations of certain eco-physiological characteristics of the broad and long-pincered crayfish in connection with their interspecific relationships. [Translation from: Lietuvas TSR Mokslu Akademijos Darbai, C Serija 2(40), 279-286, 1966.]

The results are described of eco-physiological investigations of the broad-pincered (Astacus astacus L.J.) crayfish and the long-pincered (Astacus astacus Esch.) crayfish, conducted in 1963—64 in the Institute of Zoology and Parasitology of the Academy of Science of the Lithuanian SSR, for the purpose of studying the interspecific relationship of these two species. In the course of the investigation were determined: the influence of the temperature of the environment on the consumption of oxygen by full grown individuals of both species and on the respiratory movements of the scaphognathites, the threshold temperatures and the saturation of the water by oxygen, the diel activity in the winter period.

Characteristics of the sportsmanship and unsportsmanlike conduct evaluation tools in youth sport

World Conference on Psychology and Sociology 2012