4.7 Article

Global Quantification of Glutathione S-Transferases in Human Serum Using LC-MS/MS Coupled with Affinity Enrichment

Journal

JOURNAL OF PROTEOME RESEARCH
Volume 21, Issue 5, Pages 1311-1320

Publisher

AMER CHEMICAL SOC
DOI: 10.1021/acs.jproteome.2c00049

Keywords

quantitative proteomics; GSH affinity; glutathione S-transferases; human serum; liver diseases

Funding

  1. National Key Research and Development Program of China [2017YFC0906702, 2017YFC0906703]
  2. Guangdong Basic and Applied Basic Research Foundation [2020A1515010586]
  3. Science and Technology Program of Shenzhen [JCYJ20190809144005609]

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The concentration of glutathione S-transferase (GST) in serum is considered as a potential biomarker for disease diagnosis. This study developed a new method for the enrichment and quantification of serum GSTs, which has apparent clinical potential for disease diagnosis.
The members of the glutathione S-transferase (GST) superfamily often exhibit functional overlap and can compensate for each other. Their concentrations in serum are considered as disease biomarkers. A global and quantitative evaluation of serum GSTs is therefore urgent, but there is a lack of efficient approaches due to technological limitations. GSH magnetic beads were examined for their affinity to enrich GSTs in serum, and the enriched GSTs were quantitatively targeted using a Q Exactive HF-X mass spectrometer in parallel reaction monitoring (PRM) mode. To optimize the quantification of GST peptides, sample types, trypsin digestion, and serum loading were carefully assessed; a biosynthetic method was employed to generate isotope-labeled GST peptides, and instrumental parameters were systematically optimized. A total of 134 clinical sera were collected for GST quantification from healthy donors and patients with four liver diseases. Using the new approach, GSTs in healthy sera were profiled: 14 GST peptides were quantified, and the abundance of five GST families was ranked GSTM > GSTP > GSTA > MGST1 > GSTT1, ranging from 0.1 to 4 pmol/L. Furthermore, combining the abundance of multiple GST peptides could effectively distinguish different types of liver diseases. Quantification of serum GSTs through targeted proteomics, therefore, has apparent clinical potential for disease diagnosis.

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