@Article{or.2023.030240,
AUTHOR = {XINTONG LIU, EMIKO SANADA, JIANG LI, XIAOMENG LI, HIROYUKI OSADA, NOBUMOTO WATANABE},
TITLE = {Isolation and characterization of β-transducin repeat-containing protein ligands screened using a high-throughput screening system},
JOURNAL = {Oncology Research},
VOLUME = {31},
YEAR = {2023},
NUMBER = {5},
PAGES = {645--654},
URL = {http://www.techscience.com/or/v31n5/53694},
ISSN = {1555-3906},
ABSTRACT = {β-transducin repeat-containing protein (β-TrCP) is an F-box protein subunit of the E3 Skp1-Cullin-F box (SCF) type ubiquitin-ligase complex, and provides the substrate specificity for the ligase. To find potent ligands of β-TrCP useful for the proteolysis targeting chimera (PROTAC) system using β-TrCP in the future, we developed a high-throughput screening system for small molecule β-TrCP ligands. We screened the chemical library utilizing the system and obtained several hit compounds. The effects of the hit compounds on <i>in vitro</i> ubiquitination activity of SCF<sup>β-TrCP1</sup> and on downstream signaling pathways were examined. Hit compounds NPD5943, NPL62020-01, and NPL42040-01 inhibited the TNFα-induced degradation of IκBα and its phosphorylated form. Hence, they inhibited the activation of the transcription activity of NF-κB, indicating the effective inhibition of β-TrCP by the hit compounds in cells. Next, we performed an <i>in silico</i> analysis of the hit compounds to determine the important moieties of the hit compounds. Carboxyl groups of NPL62020-01 and NPL42040-01 and hydroxyl groups of NPD5943 created hydrogen bonds with β-TrCP similar to those created by intrinsic target phosphopeptides of β-TrCP. Our findings enhance our knowledge of useful small molecule ligands of β-TrCP and the importance of residues that can be ligands of β-TrCP.},
DOI = {10.32604/or.2023.030240}
}