Preparation and characterization of vorinostat-coated beads for profiling of novel target proteins

Congcong Lu; Kai Zhang; Yi Zhang; Minjia Tan; Yijun Li; Xiwen He; Yukui Zhang

doi:10.1016/j.chroma.2014.10.098

Preparation and characterization of vorinostat-coated beads for profiling of novel target proteins

J Chromatogr A. 2014 Dec 12:1372C:34-41. doi: 10.1016/j.chroma.2014.10.098. Epub 2014 Nov 5.

Authors

Congcong Lu¹, Kai Zhang², Yi Zhang³, Minjia Tan³, Yijun Li¹, Xiwen He¹, Yukui Zhang⁴

Affiliations

¹ State Key Laboratory of Medicinal Chemical Biology & Department of Chemistry, Nankai University, Tianjin 300071, PR China.
² State Key Laboratory of Medicinal Chemical Biology & Department of Chemistry, Nankai University, Tianjin 300071, PR China; Department of Biochemistry and Molecular Biology & Tianjin Key Laboratory of Medical Epigenetics, Basic Medical School, Tianjin Medical University, Tianjin 300070, PR China. Electronic address: zhangkai@nankai.edu.cn.
³ The Chemical Proteomics Center and State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, PR China.
⁴ State Key Laboratory of Medicinal Chemical Biology & Department of Chemistry, Nankai University, Tianjin 300071, PR China; National Chromatographic Research and Analysis Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, PR China.

PMID: 25465005
DOI: 10.1016/j.chroma.2014.10.098

Abstract

Inhibitors of histone deacetylases (HDACs) have been considered to be new anticancer agents. As a key inhibitor of HDAC, vorinostat can cause growth arrest and death of a broad of transformed cells and interact with a variety of substrates. A comprehensive analysis of proteins interacting with HDAC inhibitors is of great importance in understanding molecular mechanisms of the drugs. Here, we reported the preparation and characterization of vorinostat-coated beads for profiling of novel target proteins of vorinostat (a key HDAC inhibitor). The enriched proteins were further analyzed by HPLC-MS/MS. Besides the known substrates, there were also several novel enriched protein candidates, one of which was a metalloenzyme α-enolase (ENO-1). According to our best knowledge, it is the first time that ENO-1 has been detected as a potential target of vorinostat through chemoproteomics approach. Further competition analysis indicated that ENO-1 may be co-enriched as a substrate complex. Our results demonstrated that the chemical probe combined with proteomics approach may be developed as a potential tool to identify target proteins of drugs.

Keywords: Chemical probe; HPLC–MS/MS; Proteomics; Target proteins; Vorinostat.