A multiple reaction monitoring method for absolute quantification of the human liver alcohol dehydrogenase ADH1C1 isoenzyme

Dariusz J. Janecki, Kerry G. Bemis, Tony J. Tegeler, Paresh C. Sanghani, Lanmin Zhai, Thomas Hurley, William F. Bosron, Mu Wang

Research output: Contribution to journalArticle

83 Citations (Scopus)

Abstract

Although significant progress has been made in protein quantification using mass spectrometry during recent years, absolute protein quantification in complex biological systems remains a challenging task in proteomics. The use of stable isotope-labeled standard peptide is the most commonly used strategy for absolute quantification, but it might not be suitable in all instances. Here we report an alternative strategy that employs a stable isotope-labeled intact protein as an internal standard to absolutely quantify the alcohol dehydrogenase (ADH) expression level in a human liver sample. In combination with a new targeted proteomics approach employing the method of multiple reaction monitoring (MRM), we precisely and quantitatively measured the absolute protein expression level of an ADH isoenzyme, ADH1C1, in human liver. Isotope-labeled protein standards are predicted to be particularly useful for measurement of highly homologous isoenzymes such as ADHs where multiple signature peptides can be examined by MRM in a single experiment.

Original languageEnglish
Pages (from-to)18-26
Number of pages9
JournalAnalytical Biochemistry
Volume369
Issue number1
DOIs
StatePublished - Oct 1 2007

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Alcohol Dehydrogenase
Liver
Isoenzymes
Monitoring
Isotopes
Proteins
Proteomics
Peptides
Biological systems
Mass spectrometry
Mass Spectrometry
Experiments

Keywords

  • Absolute quantification
  • Alcohol dehydrogenase
  • Human liver
  • LC-MS/MS
  • Multiple reaction monitoring

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

A multiple reaction monitoring method for absolute quantification of the human liver alcohol dehydrogenase ADH1C1 isoenzyme. / Janecki, Dariusz J.; Bemis, Kerry G.; Tegeler, Tony J.; Sanghani, Paresh C.; Zhai, Lanmin; Hurley, Thomas; Bosron, William F.; Wang, Mu.

In: Analytical Biochemistry, Vol. 369, No. 1, 01.10.2007, p. 18-26.

Research output: Contribution to journalArticle

Janecki, Dariusz J. ; Bemis, Kerry G. ; Tegeler, Tony J. ; Sanghani, Paresh C. ; Zhai, Lanmin ; Hurley, Thomas ; Bosron, William F. ; Wang, Mu. / A multiple reaction monitoring method for absolute quantification of the human liver alcohol dehydrogenase ADH1C1 isoenzyme. In: Analytical Biochemistry. 2007 ; Vol. 369, No. 1. pp. 18-26.
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