Measurement of Enzyme Isotope Effects.
| Autor: | Kholodar SA; University of Iowa, Iowa City, IA, United States., Ghosh AK; University of Iowa, Iowa City, IA, United States., Kohen A; University of Iowa, Iowa City, IA, United States. Electronic address: amnon-kohen@uiowa.edu. |
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| Jazyk: | angličtina |
| Zdroj: | Methods in enzymology [Methods Enzymol] 2017; Vol. 596, pp. 43-83. Date of Electronic Publication: 2017 Aug 02. |
| DOI: | 10.1016/bs.mie.2017.06.033 |
| Abstrakt: | Enzyme isotope effects, or the kinetic effects of "heavy" enzymes, refer to the effect of isotopically labeled protein residues on the enzyme's activity or physical properties. These effects are increasingly employed in the examination of the possible contributions of protein dynamics to enzyme catalysis. One hypothesis assumed that isotopic substitution of all 12 C, 14 N, and nonexchangeable 1 H by 13 C, 15 N, and 2 H, would slow down protein picosecond to femtosecond dynamics without any effect on the system's electrostatics following the Born-Oppenheimer approximation. It was suggested that reduced reaction rates reported for several "heavy" enzymes accords with that hypothesis. However, numerous deviations from the predictions of that hypothesis were also reported. Current studies also attempt to test the role of individual residues by site-specific labeling or by labeling a pattern of residues on activity. It appears that in several systems the protein's fast dynamics are indeed reduced in "heavy" enzymes in a way that reduces the probability of barrier crossing of its chemical step. Other observations, however, indicated that slower protein dynamics are electrostatically altered in isotopically labeled enzymes. Interestingly, these effects appear to be system dependent, thus it might be premature to suggest a general role of "heavy" enzymes' effect on catalysis. (© 2017 Elsevier Inc. All rights reserved.) |
| Databáze: | MEDLINE |
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