Online oxygen kinetic isotope effects using membrane inlet mass spectrometry can differentiate between oxidases for mechanistic studies and calculation of their contributions to oxygen consumption in whole tissues

Mun Hon Cheah; A. Harvey Millar; Ruth C. Myers; David A. Day; Justine Roth; Warwick Hillier; Murray R. Badger

doi:10.1021/ac501086n

Online oxygen kinetic isotope effects using membrane inlet mass spectrometry can differentiate between oxidases for mechanistic studies and calculation of their contributions to oxygen consumption in whole tissues

Mun Hon Cheah^*, A. Harvey Millar, Ruth C. Myers, David A. Day, Justine Roth, Warwick Hillier, Murray R. Badger

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

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Abstract

The reduction chemistry of molecular oxygen underpins the energy metabolism of multicellular organisms, liberating free energy needed to catalyze a plethora of enzymatic reactions. Measuring the isotope signatures of ¹⁶O and ¹⁸O during O₂ reduction can provide insights into both kinetic and equilibrium isotope effects. However, current methods to measure O₂ isotope signatures are time-consuming and disruptive. This paper describes the application of membrane inlet mass spectrometry to determine the oxygen isotope discrimination of a range of O ₂-consuming reactions, providing a rapid and convenient method for determining these values. A survey of oxygenase and oxidase reactions provides new insights into previously uncharacterized amino acid oxidase enzymes. Liquid and gas phase measurements show the ease of assays using this approach for purified enzymes, biological extracts and intact tissues.

Original language	English
Pages (from-to)	5171-5178
Number of pages	8
Journal	Analytical Chemistry
Volume	86
Issue number	10
DOIs	https://doi.org/10.1021/ac501086n
Publication status	Published - 20 May 2014

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Cheah, M. H., Millar, A. H., Myers, R. C., Day, D. A., Roth, J., Hillier, W., & Badger, M. R. (2014). Online oxygen kinetic isotope effects using membrane inlet mass spectrometry can differentiate between oxidases for mechanistic studies and calculation of their contributions to oxygen consumption in whole tissues. Analytical Chemistry, 86(10), 5171-5178. https://doi.org/10.1021/ac501086n

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title = "Online oxygen kinetic isotope effects using membrane inlet mass spectrometry can differentiate between oxidases for mechanistic studies and calculation of their contributions to oxygen consumption in whole tissues",

abstract = "The reduction chemistry of molecular oxygen underpins the energy metabolism of multicellular organisms, liberating free energy needed to catalyze a plethora of enzymatic reactions. Measuring the isotope signatures of 16O and 18O during O2 reduction can provide insights into both kinetic and equilibrium isotope effects. However, current methods to measure O2 isotope signatures are time-consuming and disruptive. This paper describes the application of membrane inlet mass spectrometry to determine the oxygen isotope discrimination of a range of O 2-consuming reactions, providing a rapid and convenient method for determining these values. A survey of oxygenase and oxidase reactions provides new insights into previously uncharacterized amino acid oxidase enzymes. Liquid and gas phase measurements show the ease of assays using this approach for purified enzymes, biological extracts and intact tissues.",

author = "Cheah, {Mun Hon} and Millar, {A. Harvey} and Myers, {Ruth C.} and Day, {David A.} and Justine Roth and Warwick Hillier and Badger, {Murray R.}",

year = "2014",

month = may,

day = "20",

doi = "10.1021/ac501086n",

language = "English",

volume = "86",

pages = "5171--5178",

journal = "Analytical Chemistry",

issn = "0003-2700",

publisher = "American Chemical Society",

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Cheah, MH, Millar, AH, Myers, RC, Day, DA, Roth, J, Hillier, W & Badger, MR 2014, 'Online oxygen kinetic isotope effects using membrane inlet mass spectrometry can differentiate between oxidases for mechanistic studies and calculation of their contributions to oxygen consumption in whole tissues', Analytical Chemistry, vol. 86, no. 10, pp. 5171-5178. https://doi.org/10.1021/ac501086n

Online oxygen kinetic isotope effects using membrane inlet mass spectrometry can differentiate between oxidases for mechanistic studies and calculation of their contributions to oxygen consumption in whole tissues. / Cheah, Mun Hon; Millar, A. Harvey; Myers, Ruth C. et al.
In: Analytical Chemistry, Vol. 86, No. 10, 20.05.2014, p. 5171-5178.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Online oxygen kinetic isotope effects using membrane inlet mass spectrometry can differentiate between oxidases for mechanistic studies and calculation of their contributions to oxygen consumption in whole tissues

AU - Cheah, Mun Hon

AU - Millar, A. Harvey

AU - Myers, Ruth C.

AU - Day, David A.

AU - Roth, Justine

AU - Hillier, Warwick

AU - Badger, Murray R.

PY - 2014/5/20

Y1 - 2014/5/20

N2 - The reduction chemistry of molecular oxygen underpins the energy metabolism of multicellular organisms, liberating free energy needed to catalyze a plethora of enzymatic reactions. Measuring the isotope signatures of 16O and 18O during O2 reduction can provide insights into both kinetic and equilibrium isotope effects. However, current methods to measure O2 isotope signatures are time-consuming and disruptive. This paper describes the application of membrane inlet mass spectrometry to determine the oxygen isotope discrimination of a range of O 2-consuming reactions, providing a rapid and convenient method for determining these values. A survey of oxygenase and oxidase reactions provides new insights into previously uncharacterized amino acid oxidase enzymes. Liquid and gas phase measurements show the ease of assays using this approach for purified enzymes, biological extracts and intact tissues.

AB - The reduction chemistry of molecular oxygen underpins the energy metabolism of multicellular organisms, liberating free energy needed to catalyze a plethora of enzymatic reactions. Measuring the isotope signatures of 16O and 18O during O2 reduction can provide insights into both kinetic and equilibrium isotope effects. However, current methods to measure O2 isotope signatures are time-consuming and disruptive. This paper describes the application of membrane inlet mass spectrometry to determine the oxygen isotope discrimination of a range of O 2-consuming reactions, providing a rapid and convenient method for determining these values. A survey of oxygenase and oxidase reactions provides new insights into previously uncharacterized amino acid oxidase enzymes. Liquid and gas phase measurements show the ease of assays using this approach for purified enzymes, biological extracts and intact tissues.

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DO - 10.1021/ac501086n

M3 - Article

SN - 0003-2700

VL - 86

SP - 5171

EP - 5178

JO - Analytical Chemistry

JF - Analytical Chemistry

IS - 10

ER -

Online oxygen kinetic isotope effects using membrane inlet mass spectrometry can differentiate between oxidases for mechanistic studies and calculation of their contributions to oxygen consumption in whole tissues

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