S-Nitrosoglutathione in Rat Cerebellum: Identification and Quantification by Liquid Chromatography-Mass Spectrometry

Authors

  • Ina Kluge,

    1. Brain Research Institute, University of Zürich, Zürich, Switzerland
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  • Ursula Gutteck-Amsler,

    1. Brain Research Institute, University of Zürich, Zürich, Switzerland
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    • The present address of U. Gutteck-Amsler is University Hospital, Zürich, Switzerland.

  • Markus Zollinger,

    1. Brain Research Institute, University of Zürich, Zürich, Switzerland
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    • The present address of Dr. M. Zollinger is Novartis Pharma Inc., Basel, Switzerland.

  • Kim Quang Do

    Corresponding author
    1. Brain Research Institute, University of Zürich, Zürich, Switzerland
      Address correspondence and reprint requests to Dr. K. Q. Do at Brain Research Institute, University of Zürich, August Forel-Str. 1, 8029 Zürich, Switzerland.
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Address correspondence and reprint requests to Dr. K. Q. Do at Brain Research Institute, University of Zürich, August Forel-Str. 1, 8029 Zürich, Switzerland.

Abstract

Abstract: Given the extreme lability and the facile inactivation of the messenger nitric oxide (NO) by many reactive biochemical species, it has been suggested that some intermediate compounds, for example, S-nitrosothiols, may act to stabilize NO and at the same time to preserve its biological activity. To test this hypothesis, we investigated if the S-nitrosothiol of glutathione, which is the predominant low molecular weight thiol in CNS, is present in the rat brain. The HPLC analysis of cerebellar extract from [35S]cysteine-prelabeled slices suggested that S-nitrosoglutathione (GSNO) was indeed present in rat brain. To detect endogenous GSNO, a methodology based on liquid chromatography-mass spectrometry was developed. Besides an unequivocal identification of the endogenous GSNO, this method also permitted its precise quantification using 15N-labeled GSNO ([15N]-GSNO) as internal standard. GSNO level in adult cerebellum amounts to 15.4 ± 1.4 pmol/mg of protein. This is the first direct demonstration of the presence of endogenous GSNO in CNS. The packaging of NO in the form of GSNO might serve to facilitate its transport, prolong its life, and target its delivery to specific effectors.

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