標題: Qualitative analysis of the fluorophosphonate-based chemical probes using the serine hydrolases from mouse liver and poly-3-hydroxybutyrate depolymerase (PhaZ) from Bacillus thuringiensis
作者: Huang, Yi-Long
Chung, Tsai-Wen
Chang, Chia-Mao
Chen, Chih-Hau
Liao, Chen-Chung
Tsay, Yeou-Guang
Shaw, Gwo-Chyuan
Liaw, Shwu-Huey
Sun, Chung-Ming
Lin, Chao-Hsiung
應用化學系
Department of Applied Chemistry
關鍵字: Activity-based probe;Serine hydrolase;PhaZ;Esterase;Fluorophosphonate
公開日期: 1-Nov-2012
摘要: 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 activity-based 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.
URI: http://dx.doi.org/10.1007/s00216-012-6349-0
http://hdl.handle.net/11536/20399
ISSN: 1618-2642
DOI: 10.1007/s00216-012-6349-0
期刊: ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume: 404
Issue: 8
起始頁: 2387
結束頁: 2396
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