title: Mapping sites of aspirin-induced acetylations in live cells by quantitative acid-cleavable activity-based protein profiling (QA-ABPP).
abstract: Target-identification and understanding of mechanism-of-action (MOA) are challenging for development of small-molecule probes and their application in biology and drug discovery. For example, although aspirin has been widely used for more than 100 years, its molecular targets have not been fully characterized. To cope with this challenge, we developed a novel technique called quantitative acid-cleavable activity-based protein profiling (QA-ABPP) with combination of the following two parts: (i) activity-based protein profiling (ABPP) and iTRAQ™ quantitative proteomics for identification of target proteins and (ii) acid-cleavable linker-based ABPP for identification of peptides with specific binding sites. It is known that reaction of aspirin with its target proteins leads to acetylation. We thus applied the above technique using aspirin-based probes in human cancer HCT116 cells. We identified 1110 target proteins and 2775 peptides with exact acetylation sites. By correlating these two sets of data, 523 proteins were identified as targets of aspirin. We used various biological assays to validate the effects of aspirin on inhibition of protein synthesis and induction of autophagy which were elicited from the pathway analysis of Aspirin target profile. This technique is widely applicable for target identification in the field of drug discovery and biology, especially for the covalent drugs.
authors.affiliation: Department of Biological Sciences, 14 Science Drive 4, National University of Singapore, Singapore, 117543.
authors.forename: Jigang
authors.initials: J
authors.lastname: Wang
authors.affiliation: Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Science, Baodao Xincun, Danzhou, Hainan, P.R. China, 571737.
authors.forename: Chong-Jing
authors.initials: CJ
authors.lastname: Zhang
authors.affiliation: Department of Physiology, Yong Loo Lin School of Medicine, Block MD9, 2 Medical Drive, National University of Singapore, Singapore 117597.
authors.forename: Jianbin
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Department of Anaesthesiology, Singapore General Hospital, Outram Road, Singapore, 169608.
authors.forename: Yingke
authors.initials: Y
authors.lastname: He
authors.affiliation: Department of Biological Sciences, 14 Science Drive 4, National University of Singapore, Singapore, 117543.
authors.forename: Yew Mun
authors.initials: YM
authors.lastname: Lee
authors.affiliation: Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Science, Baodao Xincun, Danzhou, Hainan, P.R. China, 571737.
authors.forename: Songbi
authors.initials: S
authors.lastname: Chen
authors.affiliation: Department of Biological Sciences, 14 Science Drive 4, National University of Singapore, Singapore, 117543.
authors.forename: Teck Kwang
authors.initials: TK
authors.lastname: Lim
authors.affiliation: Department of Physiology, Yong Loo Lin School of Medicine, Block MD9, 2 Medical Drive, National University of Singapore, Singapore 117597.
authors.forename: Shukie
authors.initials: S
authors.lastname: Ng
authors.affiliation: Department of Physiology, Yong Loo Lin School of Medicine, Block MD9, 2 Medical Drive, National University of Singapore, Singapore 117597.
authors.forename: Han-Ming
authors.initials: HM
authors.lastname: Shen
authors.affiliation: Department of Biological Sciences, 14 Science Drive 4, National University of Singapore, Singapore, 117543.
authors.forename: Qingsong
authors.initials: Q
authors.lastname: Lin
mesh_terms: D000107:Acetylation / Q000187:drug effects; D001241:Aspirin / Q000737:chemistry* / Q000378:metabolism; D001665:Binding Sites; D055808:Drug Discovery; D006801:Humans; D010455:Peptides / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D040081:Protein Array Analysis; D040901:Proteomics
chemical_list: D010455:Peptides; D001241:Aspirin