Glucose binding isotope effects in the ternary complex of brain hexokinase demonstrate partial relief of ground-state destabilization

Brett E. Lewis, Vern L. Schramm

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Abstract

Isotope effects can yield detailed information about contacts made between a bound compound and its host receptor or enzyme. In this study, we have measured isotope effects upon the equilibrium constant for the association of glucose and human brain hexokinase [E.C.2.7.1.1] in the presence of β,γ-CH2-ATP and compared these with data for the same equilibrium in the absence of ATP-analogue. We have found isotope effects of 1.012, 0.929, 1.031, 1.052, 0.998, and 1.032 for the competitive binding of [1-3H]-, [2-3H]-, [3-3H]-, [4-3H]-, [5-3H]-, or [6,6-3H2]- and either [2-14C]- or [6-14C]glucose to brain hexokinase. We observed changes only at H1, H5, and H6, and we attribute these to a slight change in the position of Asn683 and Glu742 due to nucleotide binding and to partial satisfaction of activated OH6 by the terminal nucleotide phosphorus.

Original languageEnglish (US)
Pages (from-to)4672-4673
Number of pages2
JournalJournal of the American Chemical Society
Volume125
Issue number16
DOIs
StatePublished - Apr 23 2003

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Hexokinase
Isotopes
Ground state
Glucose
Brain
Adenosinetriphosphate
Nucleotides
Adenosine Triphosphate
Competitive Binding
Equilibrium constants
Phosphorus
Enzymes
Association reactions

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

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abstract = "Isotope effects can yield detailed information about contacts made between a bound compound and its host receptor or enzyme. In this study, we have measured isotope effects upon the equilibrium constant for the association of glucose and human brain hexokinase [E.C.2.7.1.1] in the presence of β,γ-CH2-ATP and compared these with data for the same equilibrium in the absence of ATP-analogue. We have found isotope effects of 1.012, 0.929, 1.031, 1.052, 0.998, and 1.032 for the competitive binding of [1-3H]-, [2-3H]-, [3-3H]-, [4-3H]-, [5-3H]-, or [6,6-3H2]- and either [2-14C]- or [6-14C]glucose to brain hexokinase. We observed changes only at H1, H5, and H6, and we attribute these to a slight change in the position of Asn683 and Glu742 due to nucleotide binding and to partial satisfaction of activated OH6 by the terminal nucleotide phosphorus.",
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AU - Lewis, Brett E.

AU - Schramm, Vern L.

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N2 - Isotope effects can yield detailed information about contacts made between a bound compound and its host receptor or enzyme. In this study, we have measured isotope effects upon the equilibrium constant for the association of glucose and human brain hexokinase [E.C.2.7.1.1] in the presence of β,γ-CH2-ATP and compared these with data for the same equilibrium in the absence of ATP-analogue. We have found isotope effects of 1.012, 0.929, 1.031, 1.052, 0.998, and 1.032 for the competitive binding of [1-3H]-, [2-3H]-, [3-3H]-, [4-3H]-, [5-3H]-, or [6,6-3H2]- and either [2-14C]- or [6-14C]glucose to brain hexokinase. We observed changes only at H1, H5, and H6, and we attribute these to a slight change in the position of Asn683 and Glu742 due to nucleotide binding and to partial satisfaction of activated OH6 by the terminal nucleotide phosphorus.

AB - Isotope effects can yield detailed information about contacts made between a bound compound and its host receptor or enzyme. In this study, we have measured isotope effects upon the equilibrium constant for the association of glucose and human brain hexokinase [E.C.2.7.1.1] in the presence of β,γ-CH2-ATP and compared these with data for the same equilibrium in the absence of ATP-analogue. We have found isotope effects of 1.012, 0.929, 1.031, 1.052, 0.998, and 1.032 for the competitive binding of [1-3H]-, [2-3H]-, [3-3H]-, [4-3H]-, [5-3H]-, or [6,6-3H2]- and either [2-14C]- or [6-14C]glucose to brain hexokinase. We observed changes only at H1, H5, and H6, and we attribute these to a slight change in the position of Asn683 and Glu742 due to nucleotide binding and to partial satisfaction of activated OH6 by the terminal nucleotide phosphorus.

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