Preferential Control of Forward Reaction Kinetics in Hydrogels Crosslinked with Reversible Conjugate Additions

<p>Molecular substitutions were used to demonstrate preferential control over the kinetic rate constants in a poly(ethylene glycol)-based hydrogel with two different reversible thia-conjugate addition reactions. A strong electron withdrawing nitrile group on the conjugate acceptor showed a 20-fold increase in the forward rate constant over a neutral withdrawing group, while the reverse rate constant only increased 6-fold. Rheometry experiments demonstrated that the hydrogel plateau modulus was primarily dictated by reaction equilibrium, while the stress relaxation characteristics of the hydrogel were dominated by the reverse rate constant. Furthermore, the dynamic crosslinking allowed the hydrogel to rapidly and spontaneously self-heal. These results indicate that decoupling the kinetic rate constants of bond exchange allow systematic control over dynamic covalent hydrogel bulk properties, such as their adaptability, stress relaxation ability, and self-healing properties.</p>

Preferential Control of Forward Reaction Kinetics in Hydrogels Crosslinked with Reversible Conjugate Additions

<p>Molecular substitutions were used to demonstrate preferential control over the kinetic rate constants in a poly(ethylene glycol)-based hydrogel with two different reversible thia-conjugate addition reactions. A strong electron withdrawing nitrile group on the conjugate acceptor showed a 20-fold increase in the forward rate constant over a neutral withdrawing group, while the reverse rate constant only increased 6-fold. Rheometry experiments demonstrated that the hydrogel plateau modulus was primarily dictated by reaction equilibrium, while the stress relaxation characteristics of the hydrogel were dominated by the reverse rate constant. Furthermore, the dynamic crosslinking allowed the hydrogel to rapidly and spontaneously self-heal. These results indicate that decoupling the kinetic rate constants of bond exchange allow systematic control over dynamic covalent hydrogel bulk properties, such as their adaptability, stress relaxation ability, and self-healing properties.</p>

Derivable Equations and Issues Often Ignored in the Original Michaelis-Menten Mathematical Formalism

Background: There has been recent shift from the core issue of Michaelian kinetics to issues regarding various kinds of quasi-steady-state assumptions. Derivable equations with which to determine reverse rate constant for the dissociation of enzyme-substrate complex (ES) is given less attention. Objectives: The objectives of this research are: 1) to derive other equations from differential equations whose evaluation leads to MM equation and 2) quantify based on derived equations the kinetic parameters given less attention and duration of catalytic events. Methods: A major theoretical research and experimentation using Bernfeld method. Results and Discussion: The durations for ES dissociation (ESD) into free substrate, S and enzyme, E were much shorter than the duration of ESD into E and product, P in 3 minutes duration of assay with low [S]; it was the shortest and longest in 3 and 5 minutes durations respectively with high [S]. The durations of ESD into E and P was shortest in 3 minutes duration of assay with high [S]. The values of reverse rate constant, k-1 for ESD into S and E in 3 minutes duration of assay with high [S] was » the rate constant, k2 for product formation and they are much higher than in other duration of assay. Conclusion: The equations for the determination of the durations of various events, in a given catalytic cycle were derived. The various time regimes for each event and the rate constant for the dissociation of the ES can be graphically and calculationally determined as the case may be. Substrate concentration regime and duration of assay affects rate constants.

A kinetic investigation of unimolecular reactions involving trace metals at post-combustion flue gas conditions

Environmental contextUnderstanding trace metal speciation in coal combustion flue gases is imperative to the design of effective capture technologies to prevent their release into the atmosphere. Unfortunately much of the kinetics that dictate trace metal speciation are not known and the current study focuses for the first time on the kinetics for three reactions involving mercury and one involving selenium. Rate constant expressions are provided over a broad temperature range (i.e. 298–2000 K), indicative of post-combustion flue gas conditions. AbstractAb-initio methods were carried out to calculate forward and reverse rate constant data for the following reactions: Hg + Cl2 ↔ HgCl2, HgCl + Cl ↔ HgCl2, Hg + O ↔ HgO, and Se + H2 ↔ SeH2. Theoretical predictions of bond distances, vibrational frequencies and enthalpies of reaction are compared to available experimental data to determine the level of theory most appropriate for predicting kinetic parameters. The pseudopotentials ECP60MDF and RECP60VDZ were used for mercury in combination with B3LYP or QCISD(T) methods whereas the complete 6–311++G(3df,3pd) Pople basis set with the CCSD(T) method was used for selenium. Potential energy curves for each reaction were constructed and a variational approach along with RRKM theory was used to predict rate constants from 298 to 2000 K. Reactions HgCl + Cl ↔ HgCl2 and Hg + O ↔ HgO were found to have a strong negative temperature dependence, whereas the insertion reactions Hg + Cl2 ↔ HgCl2 and Se + H2 ↔ SeH2 were found to proceed very slowly with large pre-exponential factors.

Solubilization of a tamoxifen-binding protein. Assessment of its molecular mass

Recent findings point to a role of Antioestrogen-Binding Site (ABS) in some of the growth-modulatory effects of antioestrogens. In the present study, a method for the solubilization of ABS from rat uterus microsomal fractions by using 3-(3-cholamidopropyl)dimethylammonio-1-propanesulphonate (CHAPS; 20 mM) and KCl (0.4 M) is described. Decreasing the CHAPS concentration below the critical micelle concentration led to long-term stabilization of the protein. All of the membrane-bound ABS was recovered in the extract, and only one class of binding site, with a high affinity for [3H]tamoxifen (KA = 5 x 10(8) M-1) was detectable. This binding was time-dependent and reversible: at 4 degrees C, the association rate constant was ka = 7.2 x 10(4) M-1.s-1, and the reverse rate constant was kd = 1.0 x 10(-4) s-1. Solubilized ABS exhibited an affinity and specificity similar to those of the membrane-bound sites. Under disaggregating conditions, solubilized ABS had an apparent sedimentation coefficient, s20,w, of 5.2 S and a Stokes radius of 6.4 nm. From these two values, molecular masses of 160,000 Da for the detergent-ABS complex, and 110,000 for the protein moiety, were estimated. Assessment of the size of the membrane-bound ABS by a radiation inactivation technique is also described. The ‘radiation inactivation size’, corresponding to the mass of 1 mol of protein structure(s) whose associated tamoxifen-binding activity is abolished after a single hit by ionizing radiation, was estimated to be 80,000 Da.

31P-saturation-transfer nuclear-magnetic-resonance measurements of phosphocreatine turnover in guinea-pig brain slices

The technique of 31P saturation-transfer n.m.r. was used to determine the forward and the reverse rate constants of creatine phosphotransferase in superfused guinea-pig cerebral tissues in vitro. The calculated forward rate constant of 0.22 +/- 0.03s-1 compared well with a previously reported value for rat brain in vivo [Shoubridge, Briggs & Radda (1982) FEBS Lett. 140, 288-292]. The reverse rate constant was found to be 0.55 +/- 0.10s-1. 3. By using concentrations of ATP and phosphocreatine estimated previously for this superfused preparation [Cox, Morris, Feeney & Bachelard (1983) Biochem. J. 212, 365-370], forward and reverse flux rates were calculated to be 0.68 and 0.72 mumol X s-1 X g-1 respectively. The concordance of forward and reverse fluxes contrasts with the situation observed in vitro in other tissues, and suggests that the creatine phosphotransferase reaction is at equilibrium under the conditions used here. 4. Lowering the concentration of glucose in the superfusing medium from 10mM to 0.5mM had no significant effect on phosphocreatine concentration or on the forward (ATP-generating) flux through creatine phosphotransferase. The results indicate that a normal phosphocreatine content in the presence of lowered glucose availability is reflected by an unchanged turnover rate.