Qualitative analysis of the fluorophosphonate-based chemical probes using the serine hydrolases from mouse liver and poly-3-hydroxybutyrate depolymerase (PhaZ) from Bacillus thuringiensis

Yi Long Huang, Tsai Wen Chung, Chia Mao Chang, Chih Hau Chen, Chen Chung Liao, Yeou Guang Tsay, Gwo Chyuan Shaw, Shwu Huey Liaw, Chung-Ming Sun, Chao Hsiung Lin*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

The serine hydrolase family consists of more than 200 members and is one of the largest enzyme families in the human genome. Although up to 50 % of this family remains unannotated, there are increasing evidences that activities of certain serine hydrolases are associated with diseases like cancer neoplasia, invasiveness, etc. By now, several activitybased chemical probes have been developed and are applied to profile the global activity of serine hydrolases in diverse proteomes. In this study, two fluorophosphonate (FP)-based chemical probes were synthesized. Further examination of their abilities to label and pull down serine hydrolases was conducted. In addition, the poly-3- hydroxybutyrate depolymerase (PhaZ) from Bacillus thuringiensis was demonstrated as an appropriate standard serine hydrolase, which can be applied to measure the labeling ability and pull-down efficiency of FP-based probes. Furthermore, mass spectrometry (MS) was used to identify the serine residue that covalently bonded to the active probes. Finally, these FP-based probes were shown capable of establishing the serine hydrolase profiles in diverse mouse tissues; the serine hydrolases pulled down from mouse liver organ were further identified by MS. In summary, our study provides an adequate method to evaluate the reactivity of FP-based probes targeting serine hydrolases.

Original languageEnglish
Pages (from-to)2387-2396
Number of pages10
JournalAnalytical and Bioanalytical Chemistry
Volume404
Issue number8
DOIs
StatePublished - 1 Nov 2012

Keywords

  • Activity-based probe
  • Esterase
  • Fluorophosphonate
  • PhaZ
  • Serine hydrolase

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