文献类型：期刊文献

英文题名：Unambiguous Identification of -Tubulin as the Direct Cellular Target Responsible for the Cytotoxicity of Chalcone by Photoaffinity Labeling

作者：Zhou, Bo[1];Yu, Xingxin[1,2];Zhuang, Chunlin[1,3];Villalta, Peter[4];Lin, Yong[5];Lu, Junxuan[6,7];Xing, Chengguo[1]

机构：[1]Univ Minnesota, Dept Med Chem, Coll Pharm, 2231 6th St SE, Minneapolis, MN 55455 USA;[2]East China Univ Sci & Technol, Shanghai Key Lab New Drug Design, Sch Pharm, Shanghai 200237, Peoples R China;[3]Second Mil Med Univ, Res Ctr Marine Drugs, Sch Pharm, 325 Guohe Rd, Shanghai 200433, Peoples R China;[4]Univ Minnesota, Masonic Canc Ctr, 2231 6th St SE, Minneapolis, MN 55455 USA;[5]Lovelace Resp Res Inst, Mol Biol & Lung Canc Program, Albuquerque, NM 87108 USA;[6]Penn State Univ, Coll Med, Dept Pharmacol, Hershey, PA 17033 USA;[7]Penn State Univ, Coll Med, Inst Canc, Hershey, PA 17033 USA

年份：2016

卷号：11

期号：13

起止页码：1436

外文期刊名：CHEMMEDCHEM

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000380025000006)】；

基金：This research was funded through grants from the US National Institutes of Health National Cancer Institute (NCI) (R01-CA163864 to C.X.), the China Scholarship Council's PhD Abroad Training Plan (201206580001 to C.Z.), and the National Natural Science Foundation of China (81502978 to C.Z.). Mass spectrometry was performed in the Analytical Biochemistry Shared Resource at the Masonic Cancer Center, partially supported by grant P30-CA-077598 from the NCI. We thank Dr. Susith Wickramaratne for help with MS experiments and Paul Champoux for help with flow cytometry.

语种：英文

外文关键词：chalcones; chemical probes; cytotoxicity; photoaffinity labeling; tubulin

摘要：Chalcone is a simple and potentially privileged structure in medicinal chemistry with a diverse repertoire of biological activities, among which cytotoxicity is of particular interest. The sharp structure-activity relationship (SAR) for chalcone's cytotoxicity suggests structure-specific target interactions. Despite the numerous putative targets proposed, evidence for direct target interactions in cells is unavailable. In this study, guided by the sharp cytotoxic SAR, we developed a cytotoxic chalcone-based photoaffinity labeling (PAL) probe, (E)-3-(3-azidophenyl)-1-[3,5-dimethoxy-4-(prop-2-yn-1-yloxy)phenyl]-2-methylprop-2-en-1-one (C95; IC50: 0.38 +/- 0.01m), along with two structurally similar non-cytotoxic probes. These probes were used to search for the direct cellular target responsible for chalcone's cytotoxicity through intact cell-based PAL experiments, in which -tubulin was identified to specifically interact with the cytotoxic probe (i.e., C95) but not the non-cytotoxic probes. A set of phenotypical and biochemical assays further reinforced -tubulin as the cytotoxic target of chalcones. Peptide mass quantitation by mass spectrometric analysis revealed one peptide potentially labeled by C95, providing information on chalcone's binding site on -tubulin.