Binding Isotope Effects for Interrogating Enzyme–Substrate Interactions

Christopher F. Stratton, Myles B. Poulin, Vern L. Schramm

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Citations (Scopus)

Abstract

Equilibrium binding isotope effects (BIEs) report on the bond vibrational status of enzyme substrates in the Michaelis complex prior to the transition state and how they differ from the solution state. Accordingly, BIEs provide an experimental means of interrogating enzyme–substrate interactions and inform on the influence of enzyme-mediated atomic distortions in modulating substrate reactivity. In this chapter, we outline a rapid equilibrium dialysis method that our lab has used to measure BIEs for several enzyme systems. Implementation of the rapid equilibrium dialysis approach is described in the context of our recent studies on the substrate bonding environment for the human protein lysine N-methyltransferase NSD2. A summary of BIE effects provides context for the range of experimental values.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Pages1-21
Number of pages21
Volume596
DOIs
StatePublished - 2017

Publication series

NameMethods in Enzymology
Volume596
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Fingerprint

Isotopes
Dialysis
Histone-Lysine N-Methyltransferase
Substrates
Enzymes
Methyltransferases
Lysine
Proteins

Keywords

  • Analysis of binding isotope effects
  • Catalytic site distortion
  • Equilibrium dialysis
  • Ground-state stabilization
  • Inhibitor binding effects
  • Molecular distortion
  • Transition-state analysis

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Stratton, C. F., Poulin, M. B., & Schramm, V. L. (2017). Binding Isotope Effects for Interrogating Enzyme–Substrate Interactions. In Methods in Enzymology (Vol. 596, pp. 1-21). (Methods in Enzymology; Vol. 596). Academic Press Inc.. https://doi.org/10.1016/bs.mie.2017.07.019

Binding Isotope Effects for Interrogating Enzyme–Substrate Interactions. / Stratton, Christopher F.; Poulin, Myles B.; Schramm, Vern L.

Methods in Enzymology. Vol. 596 Academic Press Inc., 2017. p. 1-21 (Methods in Enzymology; Vol. 596).

Research output: Chapter in Book/Report/Conference proceedingChapter

Stratton, CF, Poulin, MB & Schramm, VL 2017, Binding Isotope Effects for Interrogating Enzyme–Substrate Interactions. in Methods in Enzymology. vol. 596, Methods in Enzymology, vol. 596, Academic Press Inc., pp. 1-21. https://doi.org/10.1016/bs.mie.2017.07.019
Stratton CF, Poulin MB, Schramm VL. Binding Isotope Effects for Interrogating Enzyme–Substrate Interactions. In Methods in Enzymology. Vol. 596. Academic Press Inc. 2017. p. 1-21. (Methods in Enzymology). https://doi.org/10.1016/bs.mie.2017.07.019
Stratton, Christopher F. ; Poulin, Myles B. ; Schramm, Vern L. / Binding Isotope Effects for Interrogating Enzyme–Substrate Interactions. Methods in Enzymology. Vol. 596 Academic Press Inc., 2017. pp. 1-21 (Methods in Enzymology).
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