Hydrogen-tritium kinetic isotope effects

TABLE 11. The primary hydrogen-tritium kinetic isotope effects found in the reactions between various alkyl radicals and tributyltin hydride and tributyltin hydride-t... 

TABLE 9. The carbon-12/carbon-14 and secondary alpha hydrogen-tritium kinetic isotope effects for the Sat2 reactions between Y-substituted /VA-dimethylanilines and Z-substituted benzyl X-substituted benzensulfonates in acetone at 35°C ... 

The mechanism of hydrogen evolution has been investigated by impedance measurements [371] and hydrogen-tritium kinetic isotope effects [375]. The effect of halides dissolved in solution has been studied [372, 376] these ions increase the overpotential in the sequence Cl- Adsorption isotherms for halides have been derived. They conform to the Temkin adsorption model with partial charge transfer. The lateral interaction between adsorbed particles has been calculated. It is higher for Br- than for I- and increases with overpotential on account of the weakening in the metal-halide bond. Thus, halides are substantial poisons for hydrogen evolution on iron. Poisons also include metal ions such as Cd2+, Zn2+, and Mn2+ [26]. 

Karsten, W.E., Hwang, C.C. and Cook, P.F. (1999). Alpha-secondary tritium kinetic isotope effects indicate hydrogen tunneling and coupled motion occur in the oxidation of L-malate by NAD-malic enzyme. Biochemistry 38, 4398-4402... 

The data given in Table 7 indicate that the deuterium-tritium kinetic isotope effect in hydrogen exchange with bases is in fact positive and lies within the limits of approximate theoretical estimates (Bigeleisen, 1949 Melander, 1960 Zollinger, 1958). The differences in the values... 

Melander first sought for a kinetic isotope effect in aromatic nitration he nitrated tritiobenzene, and several other compounds, in mixed acid and found the tritium to be replaced at the same rate as protium (table 6.1). Whilst the result shows only that the hydrogen is not appreciably loosened in the transition state of the rate-determining step, it is most easily understood in terms of the S 2 mechanism with... 

A special type of substituent effect which has proved veiy valuable in the study of reaction mechanisms is the replacement of an atom by one of its isotopes. Isotopic substitution most often involves replacing protium by deuterium (or tritium) but is applicable to nuclei other than hydrogen. The quantitative differences are largest, however, for hydrogen, because its isotopes have the largest relative mass differences. Isotopic substitution usually has no effect on the qualitative chemical reactivity of the substrate, but often has an easily measured effect on the rate at which reaction occurs. Let us consider how this modification of the rate arises. Initially, the discussion will concern primary kinetic isotope effects, those in which a bond to the isotopically substituted atom is broken in the rate-determining step. We will use C—H bonds as the specific topic of discussion, but the same concepts apply for other elements. 

Transition state theory has been useful in providing a rationale for the so-called kinetic isotope effect. The kinetic isotope effect is used by enzy-mologists to probe various aspects of mechanism. Importantly, measured kinetic isotope effects have also been used to monitor if non-classical behaviour is a feature of enzyme-catalysed hydrogen transfer reactions. The kinetic isotope effect arises because of the differential reactivity of, for example, a C-H (protium), a C-D (deuterium) and a C-T (tritium) bond. 

These reactions proceed through symmetrical transition states [H H H] and with rate constants kn,HH and kH,DH, respectively. The ratio of rate constants, kH,HH/kH,DH> defines a primary hydrogen kinetic isotope effect. More precisely it should be regarded as a primary deuterium kinetic isotope effect because for hydrogen there is also the possibility of a tritium isotope effect. The term primary indicates that bonds at the site of isotopic substitution the isotopic atom are being made or broken in the course of reaction. Within the limits of TST such isotope effects are typically in the range of 4 to 8 (i.e. 4 < kH,HH/kH,DH < 8). 

We will use reaction 10.15 to illustrate two important concepts of kinetic isotope effect studies. The first concerns the relation between isotope effects of different isotopes of the same element, say D and T. We denote the rate constant of reaction 10.15 by kn and consider isotope effects when one hydrogen in the a-position is substituted by deuterium or tritium ... 

Timelines can also be portrayed in charts or figures, as illustrated in excerpt 14D. In fact, charts and figures represent excellent ways to illustrate how smaller, individual projects contribute to larger research goals and how smaller projects complement one another and overlap in time. (Note In excerpt 14D, Kohen uses the following abbreviations in his chart, each defined previously in the proposal hydrogen (H), tritium (T), deuterium (D), kinetic isotope effect (KIE), dihydrofolate reductase (a relatively small protein) (DHFR), and wild type (WT).)... 

The second substrate glyoxylate approaches from the other side of the molecule and condenses as is shown. Since any one of the three protons in either R or S chiral acetyl-CoA might have been abstracted by base B, several possible combinations of isotopes are possible in the L-malate formed. One of the results of the experiment using chiral (R) acetyl-CoA is illustrated in Eq. 13-43. The reader can easily tabulate the results of removal of the 2H or 3H. However, notice that if the base -B removes 2H (D) or 3H (T) the reaction will be much slower because of the kinetic isotope effects which are expected to be Hk/"k 7 and "k/ k = 16. A second important fact is that the pro-R hydrogen at C-3 in malate is specifically exchanged out into water by the action of fumarate hydratase. From the distribution of tritium in the malate and fumarate formed using the two chiral acetates, the inversion by malate synthase was established. See Kyte231 for a detailed discussion. 

Bryce-Smith et al. (1954) have established the presence of a positive kinetic isotope effect in metallation reactions. For example, ordinary toluene is metallated by ethyl potassium five times faster than is C6H5. CD3. Gronowitz and Halvarson (1955) have found that thiophene of ordinary isotopic composition is metallated six times faster than thiophene in which the hydrogen has been replaced by tritium. 

Feeding experiments with doubly labeled 3-hydroxy-4-[14C]methoxy-A-meth-yl-(f -[3H]- and -(5)-[3H]A,-benzylamines in King Alfred daffodils produced oduline (186) with high (82-85%) tritium retention (139). This observation suggested that the incorporation of A-methylisovanillamine into 186 occurred by a nonstereospecific process in which hydrogen removal from the benzylic position was governed by a kinetic isotope effect. 

---