title: The small molecule '1-(4-biphenylylcarbonyl)-4-(5-bromo-2-methoxybenzyl) piperazine oxalate' and its derivatives regulate global protein synthesis by inactivating eukaryotic translation initiation factor 2-alpha.
abstract: By environmental stresses, cells can initiate a signaling pathway in which eukaryotic translation initiation factor 2-alpha (eIF2-α) is involved to regulate the response. Phosphorylation of eIF2-α results in the reduction of overall protein neogenesis, which allows cells to conserve resources and to reprogram energy usage for effective stress control. To investigate the role of eIF2-α in cell stress responses, we conducted a viability-based compound screen under endoplasmic reticulum (ER) stress condition, and identified 1-(4-biphenylylcarbonyl)-4-(5-bromo-2-methoxybenzyl) piperazine oxalate (AMC-01) and its derivatives as eIF2-α-inactivating chemical. Molecular characterization of this signaling pathway revealed that AMC-01 induced inactivation of eIF2-α by phosphorylating serine residue 51 in a dose- and time-dependent manner, while the negative control compounds did not affect eIF2-α phosphorylation. In contrast with ER stress induction by thapsigargin, phosphorylation of eIF2-α persisted for the duration of incubation with AMC-01. By pathway analysis, AMC-01 clearly induced the activation of protein kinase RNA-activated (PKR) kinase and nuclear factor-κB (NF-κB), whereas it did not modulate the activity of PERK or heme-regulated inhibitor (HRI). Finally, we could detect a lower protein translation rate in cells incubated with AMC-01, establishing AMC-01 as a potent chemical probe that can regulate eIF2-α activity. We suggest from these data that AMC-01 and its derivative compounds can be used as chemical probes in future studies of the role of eIF2-α in protein synthesis-related cell physiology.
authors.affiliation: Department of Convergence Medicine, Asan Institute for Life Sciences, Asan Medical Center, Convergence Medicine Research Building, 43 gil Olympicro, Pungnapdong, Songpagu, Seoul, 138-736, Republic of Korea.
authors.forename: Mi-Na
authors.initials: MN
authors.lastname: Hong
authors.affiliation: Department of Convergence Medicine, Asan Institute for Life Sciences, Asan Medical Center, Convergence Medicine Research Building, 43 gil Olympicro, Pungnapdong, Songpagu, Seoul, 138-736, Republic of Korea.
authors.forename: Ky-Youb
authors.initials: KY
authors.lastname: Nam
authors.affiliation: Department of Convergence Medicine, Asan Institute for Life Sciences, Asan Medical Center, Convergence Medicine Research Building, 43 gil Olympicro, Pungnapdong, Songpagu, Seoul, 138-736, Republic of Korea.|Department of Convergence Medicine, College of Medicine, University of Ulsan, Seoul, Republic of Korea.
authors.forename: Kyung Kon
authors.initials: KK
authors.lastname: Kim
authors.affiliation: Department of Convergence Medicine, Asan Institute for Life Sciences, Asan Medical Center, Convergence Medicine Research Building, 43 gil Olympicro, Pungnapdong, Songpagu, Seoul, 138-736, Republic of Korea.
authors.forename: So-Young
authors.initials: SY
authors.lastname: Kim
authors.affiliation: Department of Convergence Medicine, Asan Institute for Life Sciences, Asan Medical Center, Convergence Medicine Research Building, 43 gil Olympicro, Pungnapdong, Songpagu, Seoul, 138-736, Republic of Korea. ik.kim@amc.seoul.kr.|Department of Convergence Medicine, College of Medicine, University of Ulsan, Seoul, Republic of Korea. ik.kim@amc.seoul.kr.
authors.forename: InKi
authors.initials: I
authors.lastname: Kim
mesh_terms: D000818:Animals; D002460:Cell Line; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000235:genetics; D059865:Endoplasmic Reticulum Stress / Q000187:drug effects / Q000235:genetics*; D005786:Gene Expression Regulation / Q000187:drug effects; D016328:NF-kappa B / Q000096:biosynthesis / Q000235:genetics; D010070:Oxalates / Q000494:pharmacology; D010879:Piperazines / Q000494:pharmacology; D014176:Protein Biosynthesis / Q000187:drug effects / Q000235:genetics*; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects; D013312:Stress, Physiological / Q000187:drug effects / Q000235:genetics*; D019284:Thapsigargin / Q000494:pharmacology; D019892:eIF-2 Kinase / Q000235:genetics* / Q000378:metabolism
chemical_list: D016328:NF-kappa B; D010070:Oxalates; D010879:Piperazines; D019284:Thapsigargin; D019892:eIF-2 Kinase
keywords: Cell stress response; Eukaryotic translation initiation factor 2-alpha; Molecular probe