A kinetic model is provided to obtain reaction rate constants in successive enzymatic reactions that are monitored using NMR spectroscopy and hyperpolarized substrates. The model was applied for simulation and analysis of the successive oxidation of choline to betaine aldehyde, and further to betaine, by the enzyme choline oxidase. This enzymatic reaction was investigated under two different sets of conditions: two different choline molecular probes were used, [1,1,2,2-D4 , 1-(13) C]choline chloride and [1,1,2,2-D4 , 2-(13) C]choline chloride, in different MR systems (clinical scanner and high-resolution spectrometer), as well as in different reactors and reaction volumes (4.8 and 0.7 mL). The kinetic analysis according to the model yielded similar results in both set-ups, supporting the robustness of the model. This was achieved despite the complex and negating influences of reaction kinetics and polarization decay, and in the presence of uncontrolled mixing characteristics, which may introduce uncertainties in both effective timing and effective pulses. The ability to quantify rate constants using hyperpolarized MR in the first seconds of consecutive enzyme activity is important for further development of the utilization of dynamic nuclear polarization-MR for biological determinations.
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[NMR paper] Weak Intermolecular Hydrogen Bonds with Fluorine: Detection and Implications for Enzymatic/Chemical Reactions, Chemical Properties, and Ligand/Protein Fluorine NMR Screening.
Weak Intermolecular Hydrogen Bonds with Fluorine: Detection and Implications for Enzymatic/Chemical Reactions, Chemical Properties, and Ligand/Protein Fluorine NMR Screening.
Related Articles Weak Intermolecular Hydrogen Bonds with Fluorine: Detection and Implications for Enzymatic/Chemical Reactions, Chemical Properties, and Ligand/Protein Fluorine NMR Screening.
Chemistry. 2016 Apr 26;
Authors: Dalvit C, Vulpetti A
Abstract
It is known that strong hydrogen-bonding interactions play an important role in many chemical and...
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04-27-2016 01:51 PM
TheProline Enamine Formation Pathway Revisited inDimethyl Sulfoxide: Rate Constants Determined via NMR
TheProline Enamine Formation Pathway Revisited inDimethyl Sulfoxide: Rate Constants Determined via NMR
Michael H. Haindl, Johnny Hioe and Ruth M. Gschwind
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/jacs.5b03420/20151006/images/medium/ja-2015-03420y_0008.gif
Journal of the American Chemical Society
DOI: 10.1021/jacs.5b03420
http://feeds.feedburner.com/~ff/acs/jacsat?d=yIl2AUoC8zA
http://feeds.feedburner.com/~r/acs/jacsat/~4/FFtVCyXk5zk
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10-06-2015 10:39 PM
Apparent rate constant mapping using hyperpolarized [1–13C]pyruvate
From The DNP-NMR Blog:
Apparent rate constant mapping using hyperpolarized pyruvate
Khegai, O., et al., Apparent rate constant mapping using hyperpolarized pyruvate. NMR in Biomedicine, 2014. 27(10): p. 1256-1265.
http://dx.doi.org/10.1002/nbm.3174
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01-26-2015 03:48 PM
Long-Lived States of Magnetically Equivalent Spins Populated by Dissolution-DNP and Revealed by Enzymatic Reactions
From The DNP-NMR Blog:
Long-Lived States of Magnetically Equivalent Spins Populated by Dissolution-DNP and Revealed by Enzymatic Reactions
Bornet, A., et al., Long-Lived States of Magnetically Equivalent Spins Populated by Dissolution-DNP and Revealed by Enzymatic Reactions. Chemistry, 2014. 20(51): p. 17113-8.
http://www.ncbi.nlm.nih.gov/pubmed/25346515
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12-17-2014 09:43 PM
Detection of Living Anionic Species in Polymerization Reactions Using Hyperpolarized NMR
From The DNP-NMR Blog:
Detection of Living Anionic Species in Polymerization Reactions Using Hyperpolarized NMR
Lee, Y., et al., Detection of Living Anionic Species in Polymerization Reactions Using Hyperpolarized NMR. J. Am. Chem. Soc., 2013. 135(12): p. 4636-4639.
http://dx.doi.org/10.1021/ja4001008
Intermediates during the anionic polymerization of styrene were observed using hyperpolarized NMR. Dissolution dynamic nuclear polarization (DNP) of monomers provides a sufficient signal-to-noise ratio for detection of 13C NMR signals in real time as the reaction progresses. Because...
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04-29-2013 06:02 PM
Detection of Living Anionic Species in Polymerization Reactions Using Hyperpolarized NMR
Detection of Living Anionic Species in Polymerization Reactions Using Hyperpolarized NMR
Youngbok Lee, Gyu Seong Heo, Haifeng Zeng, Karen L. Wooley and Christian Hilty
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/ja4001008/aop/images/medium/ja-2013-001008_0005.gif
Journal of the American Chemical Society
DOI: 10.1021/ja4001008
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http://feeds.feedburner.com/~r/acs/jacsat/~4/mn-OxDYOupY
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03-13-2013 08:55 PM
Measurement of rate constants for homodimer subunit exchange using double electronâ??electron resonance and paramagnetic relaxation enhancements
Measurement of rate constants for homodimer subunit exchange using double electronâ??electron resonance and paramagnetic relaxation enhancements
Abstract Here, we report novel methods to measure rate constants for homodimer subunit exchange using double electronâ??electron resonance (DEER) electron paramagnetic resonance spectroscopy measurements and nuclear magnetic resonance spectroscopy based paramagnetic relaxation enhancement (PRE) measurements. The techniques were demonstrated using the homodimeric protein Dsy0195 from the strictly anaerobic bacterium Desulfitobacterium...
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11-29-2012 03:14 AM
A study on the influence of fast amide exchange on the accuracy of 15N relaxation rate constants
A study on the influence of fast amide exchange on the accuracy of 15N relaxation rate constants
Abstract 15N relaxation rates of amide moieties provide insight both into global as well as local backbone dynamics of peptides and proteins. As the differences in the relaxation rates in general are small, their accurate determination is of prime importance. One potential source of error is fast amide exchange. It is well known that in its presence the effects of saturation transfer and H/D exchange may result in erroneous apparent relaxation rates R 1 and R 2. Here, the extent of...