Quantification of mitochondrial S-Nitrosylation by CysTMT6 switch assay

Christopher I. Murray, Hea Seung Chung, Helge Uhrigshardt, Jennifer E. Van Eyk

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

S-nitrosylation (SNO) is an important oxidative posttranslational modification in the regulation of cardiac mitochondria. SNO modification of several mitochondrial proteins has been associated with cardiac preconditioning and improved cell survival following ischemia/reperfusion injury. Due to their labile nature, SNO modifications are challenging to study using traditional biochemical techniques; particularly, the identification of individual modified cysteine residues. Here, we describe the details of the cysTMT 6 switch assay, a variation of the classic biotin switch protocol. The cysTMT 6 reagent provides a simplified and powerful approach to SNO detection by combining unambiguous identification of the modified cysteine residue and relative quantification of up to six samples by mass spectrometry analysis.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages169-179
Number of pages11
Volume1005
ISBN (Print)9781627033855
DOIs
StatePublished - 2013

Publication series

NameMethods in Molecular Biology
Volume1005
ISSN (Print)10643745

Fingerprint

Cysteine
Mitochondrial Proteins
Post Translational Protein Processing
Biotin
Reperfusion Injury
Mass Spectrometry
Cell Survival
Mitochondria

Keywords

  • Biotin switch assay
  • Cysteine-reactive tandem mass tags
  • Mitochondria
  • Oxidative modification
  • Quantitation
  • S-nitrosylation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Medicine(all)

Cite this

Murray, C. I., Chung, H. S., Uhrigshardt, H., & Van Eyk, J. E. (2013). Quantification of mitochondrial S-Nitrosylation by CysTMT6 switch assay. In Methods in Molecular Biology (Vol. 1005, pp. 169-179). (Methods in Molecular Biology; Vol. 1005). Humana Press Inc.. https://doi.org/10.1007/978-1-62703-386-2_14

Quantification of mitochondrial S-Nitrosylation by CysTMT6 switch assay. / Murray, Christopher I.; Chung, Hea Seung; Uhrigshardt, Helge; Van Eyk, Jennifer E.

Methods in Molecular Biology. Vol. 1005 Humana Press Inc., 2013. p. 169-179 (Methods in Molecular Biology; Vol. 1005).

Research output: Chapter in Book/Report/Conference proceedingChapter

Murray, CI, Chung, HS, Uhrigshardt, H & Van Eyk, JE 2013, Quantification of mitochondrial S-Nitrosylation by CysTMT6 switch assay. in Methods in Molecular Biology. vol. 1005, Methods in Molecular Biology, vol. 1005, Humana Press Inc., pp. 169-179. https://doi.org/10.1007/978-1-62703-386-2_14
Murray CI, Chung HS, Uhrigshardt H, Van Eyk JE. Quantification of mitochondrial S-Nitrosylation by CysTMT6 switch assay. In Methods in Molecular Biology. Vol. 1005. Humana Press Inc. 2013. p. 169-179. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-62703-386-2_14
Murray, Christopher I. ; Chung, Hea Seung ; Uhrigshardt, Helge ; Van Eyk, Jennifer E. / Quantification of mitochondrial S-Nitrosylation by CysTMT6 switch assay. Methods in Molecular Biology. Vol. 1005 Humana Press Inc., 2013. pp. 169-179 (Methods in Molecular Biology).
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