scholarly journals The dependence of DNase I activity on the conformation of oligodeoxynucleotides

1997 ◽  
Vol 321 (2) ◽  
pp. 481-486 ◽  
Author(s):  
Deborah H. SUTTON ◽  
Graeme L. CONN ◽  
Tom BROWN ◽  
Andrew N. LANE

We have developed a sensitive continuous assay for nucleases using proton release. The assay has been applied to the determination of the kinetics of DNase I acting on short, defined deoxyoligonucleotides. The dependence of kcat/Km on sequence and structure of short oligonucleotide substrates has been measured: increasing lengths of AnTn sequences decrease the rate of cleavage. GƃA mismatches in which the bases pair using imino protons are cleaved quite effectively by DNase I. In contrast, tandem GƃA mismatches which use amino pairing and have BII phosphodiesters, are refractory to DNase I. Also, the DNA strands of DNAƃRNA hybrid duplexes are not cleaved by DNase I. These results show that the global conformation of a duplex and the details of its minor groove affect the cleavage efficiency by DNase I. The assay has also been used to measure the inhibition constant of the minor-groove-binding ligand propamidine. A value of 3 ƁM has been determined for binding to the sequence d(CGCGAATTCGCG)2, showing that dissociation constants can be determined even when there are no convenient optical signals for titrations.

2015 ◽  
Vol 1765 ◽  
pp. 127-132 ◽  
Author(s):  
Emmanuel P. R. Lima ◽  
Pedro C. de Lima ◽  
Marcelo Nava

ABSTRACTThe non-ferrous shape memory alloys have, normally, two problems that hinder its use in industrial scale: the natural aging and grains growth. The first degrades the memory effect, while the second, observed during the processing of alloy, modifies the temperatures which the transformations occur. Thus, the study of kynetic of recrystallization is important for enabling the control of hardened state in function of treatment time, without causing excessive grain growth. Therefore, the objective of this study is to determine the kinetics of recrystallization of Cu-14Al-4Ni shape memory alloy, based on an empirical law of the formation of Jonhson-Mehl-Avrami, as well as their activation energies for grain growth process according to the empirical Arrhenius law. The alloy was vacuum melted in an induction furnace. After casting, the bulk samples of the alloy were homogenized for 24 hours, solubilized and hot rolled followed by water-quenching to initiate the recrystallization. Then, different samples were annealed at temperatures close to the peak, start and end of the DSC curve. Following the heat treatments, the samples were submitted to mechanical tests and the values of the properties were correlated to the fraction transformed for determination of recrystallization’s kinetic. For the characterization of the grain growth process, analyses in optical microscopy were accomplished and all annealed samples were examined by statistical metallography and the grain sizes were measured. After measurements, the ln[-ln(1-Yrec)] x ln(t) and the ln [D-Do] x 1/T diagrams were plotted to determine the parameters of Jonhson-Mehl-Avrami equation and the activation energy of the process, respectively. The results showed that the equation of the recrystallized fraction follows the empiric law of the formation of Jonhson-Mehl-Avrami for the considered property, as well as, also showed that the alloy Cu-14Al-4Ni is extremely sensitive to temperature variation in which the alloy is treated, having a dual kinetics of grain growth. In the first domain, between 670 and 710°C, the diagram provides a value for the activation energy equal to 39.32 KJ/mol, in the second domain, between 710 and 790°C, the diagram provides a value for the activation energy equal to 9.01 KJ/mol.


1986 ◽  
Vol 234 (3) ◽  
pp. 705-715 ◽  
Author(s):  
N Rhodes ◽  
C N Morris ◽  
S Ainsworth ◽  
J Kinderlerer

The kinetics of pyruvate kinase from Saccharomyces cerevisiae were studied at 25 degrees C and pH 6.2 as a function of the concentrations of ADP, phosphoenolpyruvate, Mg2+ and either NH4+ or K+. The data were analysed by the exponential model for four substrates, obtained by extension of the model described by Ainsworth, Kinderlerer & Gregory [(1983) Biochem. J. 209, 401-411]. On that basis, it was concluded that NH4+ binding is almost non-interactive but leads to the appearance of positive interaction in the velocity response to increase in its concentration because of positive interactions with phosphoenolpyruvate and Mg2+. The data obtained with K+ lead to the same conclusions and differ only in suggesting that NH4+ is bound more strongly to the enzyme than is K+. Both data sets are used as the basis for a discussion of the substrate interactions of pyruvate kinase and it appears therefrom that the heterotropic interactions accord with what is known of the events that take place at the active site during catalysis. The paper also reports a determination of the dissociation constants for the NH4+ complexes with ADP and phosphoenolpyruvate and an examination of the simultaneous activation of pyruvate kinase by K+ and NH4+ ions.


ADMET & DMPK ◽  
2020 ◽  
Vol 9 (1) ◽  
pp. 57-68
Author(s):  
Alisa Elezović ◽  
Amina Marić ◽  
Amila Biščević ◽  
Jasmina Hadžiabdić ◽  
Selma Škrbo ◽  
...  

The kinetics of passive transport of ketoprofen and metformin, as model substances for high and low permeability, respectively, across the artificial membrane under the influence of the pH of donor solution was investigated. There was an upward trend in the apparent permeation coefficient (Papp) of ketoprofen with the decrease in pH to a value close to pKa. At the pH value below pKa the permeation coefficient had lower value, due to the higher retention of ketoprofen in the artificial membrane. Metformin is a low permeable compound, and the highest permeation values were recorded at pH 7.4. Two dissociation constants determine that metformin at physiological pH exists as a hydrophilic cationic molecule, i.e. predominantly in ionized form. At pH values below 2.8, metformin mainly exists in diprotonated form, and it was, thus, very poorly permeable. The highest retention, i.e. affinity of both ketoprofen and metformin to the membrane, was at the lowest pH values, which is explained by different mechanisms. At higher pH values of the donor compartment, the substances showed significantly less affinity to the membrane. The obtained values of apparent permeation coefficients at studied pH values showed a good correlation with the obtained experimental values by other in vitro methods.    


2014 ◽  
Vol 84 (Supplement 1) ◽  
pp. 25-29 ◽  
Author(s):  
Guangwen Tang

Humans need vitamin A and obtain essential vitamin A by conversion of plant foods rich in provitamin A and/or absorption of preformed vitamin A from foods of animal origin. The determination of the vitamin A value of plant foods rich in provitamin A is important but has challenges. The aim of this paper is to review the progress over last 80 years following the discovery on the conversion of β-carotene to vitamin A and the various techniques including stable isotope technologies that have been developed to determine vitamin A values of plant provitamin A (mainly β-carotene). These include applications from using radioactive β-carotene and vitamin A, depletion-repletion with vitamin A and β-carotene, and measuring postprandial chylomicron fractions after feeding a β-carotene rich diet, to using stable isotopes as tracers to follow the absorption and conversion of plant food provitamin A carotenoids (mainly β-carotene) in humans. These approaches have greatly promoted our understanding of the absorption and conversion of β-carotene to vitamin A. Stable isotope labeled plant foods are useful for determining the overall bioavailability of provitamin A carotenoids from specific foods. Locally obtained plant foods can provide vitamin A and prevent deficiency of vitamin A, a remaining worldwide concern.


2014 ◽  
Vol 2 (42) ◽  
pp. 263-263
Author(s):  
Farhoush Kiani ◽  
Mahmoud Tajbakhsh ◽  
Fereydoon Ashrafi ◽  
Nesa Shafiei ◽  
Azar Bahadori ◽  
...  

2020 ◽  
Author(s):  
Camilo A. Mesa ◽  
Ludmilla Steier ◽  
Benjamin Moss ◽  
Laia Francàs ◽  
James E. Thorne ◽  
...  

<p><i>Operando</i> spectroelectrochemical analysis is used to determine the water oxidation reaction kinetics for hematite photoanodes prepared using four different synthetic procedures. Whilst these photoanodes exhibit very different current / voltage performance, their underlying water oxidation kinetics are found to be almost invariant. Lower photoanode performance was found to correlate with the observation of optical signals indicative of charge accumulation in mid-gap oxygen vacancy states, indicating these states do not contribute directly to water oxidation.</p>


2014 ◽  
pp. 228-231 ◽  
Author(s):  
Maciej Wojtczak ◽  
Aneta Antczak-Chrobot ◽  
Edyta Chmal-Fudali ◽  
Agnieszka Papiewska

The aim of the study is to evaluate the kinetics of the synthesis of dextran and other bacterial metabolites as markers of microbiological contamination of sugar beet.


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