The Mode of Action of Recombinant Mycobacterium tuberculosis Shikimate Kinase: Kinetics and Thermodynamics Analyses

Tuberculosis remains as one of the main cause of mortality worldwide due to a single infectious agent, Mycobacterium tuberculosis. The aroK-encoded M. tuberculosis Shikimate Kinase (MtSK), shown to be essential for survival of bacilli, catalyzes the phosphoryl transfer from ATP to the carbon-3 hydroxyl group of shikimate (SKH), yielding shikimate-3-phosphate and ADP. Here we present purification to homogeneity, and oligomeric state determination of recombinant MtSK. Biochemical and biophysical data suggest that the chemical reaction catalyzed by monomeric MtSK follows a rapid-equilibrium random order of substrate binding, and ordered product release. Isothermal titration calorimetry (ITC) for binding of ligands to MtSK provided thermodynamic signatures of non-covalent interactions to each process. A comparison of steady-state kinetics parameters and equilibrium dissociation constant value determined by ITC showed that ATP binding does not increase the affinity of MtSK for SKH. We suggest that MtSK would more appropriately be described as an aroL-encoded type II shikimate kinase. Our manuscript also gives thermodynamic description of SKH binding to MtSK and data for the number of protons exchanged during this bimolecular interaction. The negative value for the change in constant pressure heat capacity (ΔCp) and molecular homology model building suggest a pronounced contribution of desolvation of non-polar groups upon binary complex formation. Thermodynamic parameters were deconvoluted into hydrophobic and vibrational contributions upon MtSK:SKH binary complex formation. Data for the number of protons exchanged during this bimolecular interaction are interpreted in light of a structural model to try to propose the likely amino acid side chains that are the proton donors to bulk solvent following MtSK:SKH complex formation. © 2013 Rosado et al.

Structure and kinetics of formation of APTS/GPTS-derived organic/inorganic hybrids

The structure and the kinetics of formation of APTS/GPTS-derived organic/inorganic hybrids were studied in situ by small-angle-X-ray scattering. The data were interpreted in terms of a process of primary particles formation and growth of mass-fractal clusters. At the very early stage, the population of the nonfractal primary particles (D = 3) increases with time. As the mass-fractal clusters appear (D < 3) as a result of the aggregation process, the radius of gyration, Rg, of the clusters increases on average. At advanced stages of aggregation, the clusters grow in a mechanism in which the number of particles per clusters increases while the number of clusters diminishes with time, in such a way that the correlation volume of the clusters, Vc, fulfills the relationship Vc â̂ R g D, in agreement with a mass-fractal character of the clusters. These results supporting a cluster-cluster aggregation process, together with the typically very low value found for the mass-fractal dimension D, are in favor of a diffusion-controlled cluster aggregation mechanism. © 2013 American Chemical Society.

Kinetics of epsilon antitoxin antibodies in different strategies for active immunization of lambs against enterotoxaemia

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The Positive Effects of Priming Exercise on Oxygen Uptake Kinetics and High-Intensity Exercise Performance Are Not Magnified by a Fast-Start Pacing Strategy in Trained Cyclists

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

A Facile Measurement of Heterogeneous Electron Transfer Kinetics

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The effects of genistein and daidzein on cell proliferation kinetics in HT29 colon cancer cells: the expression of CTNNBIP1 (beta-catenin), APC (adenomatous polyposis coli) and BIRC5 (survivin)

Soybean isoflavonoids have received significant attention due to their potential anticarcinogenic and antiproliferative effects and possible role in many signal transduction pathways. However, their mechanisms of action and their molecular targets remain to be further elucidated. In this paper, we demonstrated that two soybean isoflavones (genistein and daidzein) reduced the proliferation of the human colon adenocarcinoma grade II cell line (HT-29) at concentrations of 25 and 50-100 mu M, respectively. We then investigated the effects of genistein and daidzein by RT-PCR on molecules that involved in tumor development and progression by their regulation of cell proliferation. At a concentration of 50 mu M genistein, there was suppressed expression of beta-catenin (CTNNBIP1). Neither genistein nor daidzein affected APC (adenomatous polyposis coli) or survivin (BIRC5) expression when cells were treated with concentrations of 10 or 50 mu M. These data suggest that the down-regulation of beta-catenin by genistein may constitute an important determinant of the suppression of HT-29 cell growth and may be exploited for the prevention and treatment of colon cancer.

Drying kinetics and quality of natural coffee

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Chemical Composition, Rumen Fermentation Kinetics, Digestibility and Energy Value of Cassava Leaves Hay at Different Storage Times

Cassava leaves have been widely used as a protein source for ruminants in the tropics. However, these leaves contain high level of hydro-cyanic acid (HCN) and condensed tannins (CT). There are evidences that making hay can eliminate more than 90% of HCN and that long-term storage can reduce CT levels. A complete randomized design with four replicates was conducted to determine the effect of different storage times (0-control, 60, 90 and 120 days) on chemical composition, in vitro rumen fermentation kinetics, digestibility and energy value of cassava leaves hay. Treatments were compared by analyzing variables using the GLM procedure (SAS 9.1, SAS Institute, Inc., Cary, NC). Crude protein (CP) and ether extract (EE) of the cassava hay were not affected (P > 0.05) by storage time (17.7% and 3.0%, respectively). Neutral detergent fiber, acid detergent fiber, total carbohydrate and non-fiber carbohydrate were not affected either (P>0.05) by storage time (47.5, 32.6, 72.3 and 25.8% respectively). However, other parameters were influenced. CT was lower (P<0.05) in hay after 120 days of storage compared with control (1.75% versus 3.75%, respectively). Lignin and insoluble nitrogen in neutral detergent, analyzed without sodium sulfite, were higher (P<0.01) after 120 days of storage, compared with the control (11.22 versus 13.57 and 1.65 versus 3.81% respectively). This suggests that the CT has bound to the fiber or CP and became inactive. Consequently, the in vitro digestibility of organic matter (50.36%), total digestible nutrients (44.79%) and energy (1.61 Mcal/KgMS), obtained from gas production data at 72 h of incubation, has increased (P<0.05) with storage times (56.83%, 51.53% and 1.86 Mcal/KgMS, respectively). The chemical composition and fermentative characteristics of cassava hay suffered variations during the storage period. The best values were obtained after 90 days of storage. This is probably due to the reduction in condensed tannins.

Cell cycle kinetics, apoptosis rates, DNA damage and TP53 gene expression in bladder cancer cells treated with allyl isothiocyanate (mustard essential oil)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of calcium on the osmotic dehydration kinetics and quality of pineapple

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Evaluation of Decomposition Kinetics of Poly (Ether-Ether-Ketone) by Thermogravimetric Analysis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Corrosion kinetics and topography analysis of Ti-6Al-4V alloy subjected to different mouthwash solutions

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Pluronics F-127/L-81 Binary Hydrogels as Drug-Delivery Systems: Influence of Physicochemical Aspects on Release Kinetics and Cytotoxicity

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Kinetics of rabies antibodies as a strategy for canine active immunization

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Dinitrosyl iron complexes with cysteine. kinetics studies of the formation and reactions of dnics in aqueous solution

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)