title: Polyproline-rod approach to isolating protein targets of bioactive small molecules: isolation of a new target of indomethacin.
abstract: Identification of protein targets of bioactive small molecules has been a technical hurdle of chemical genetics. Here we report a polyproline-rod approach to isolating protein targets of small molecules from cell lysates. The results indicate that insertion of a long, rigid polyproline helix between a small-molecule bait and a biotin tag boosts the capacity of affinity purification and thereby permits isolation of low-abundance or low-affinity proteins. In the course of the proof-of-concept experiments, we isolated glyoxalase 1 (GLO1) as a new target of indomethacin, a widely used antiinflammatory drug. Molecular biological experiments suggest that inhibition of GLO1 enzyme activity is related to the clinically recognized beneficial side effects of the indomethacin family of nonsteroidal antiinflammatory drugs.
authors.affiliation: The Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas 77030, USA.
authors.forename: Shin-Ichi
authors.initials: S
authors.lastname: Sato
authors.forename: Youngjoo
authors.initials: Y
authors.lastname: Kwon
authors.forename: Shinji
authors.initials: S
authors.lastname: Kamisuki
authors.forename: Neeta
authors.initials: N
authors.lastname: Srivastava
authors.forename: Qian
authors.initials: Q
authors.lastname: Mao
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Kawazoe
authors.forename: Motonari
authors.initials: M
authors.lastname: Uesugi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000378:metabolism* / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000378:metabolism; D002457:Cell Extracts / Q000737:chemistry; D002478:Cells, Cultured; D006801:Humans; D007213:Indomethacin / Q000378:metabolism* / Q000494:pharmacology; D007791:Lactoylglutathione Lyase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry*; D011506:Proteins / Q000302:isolation & purification* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000494:pharmacology
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D000970:Antineoplastic Agents; D002457:Cell Extracts; D010455:Peptides; D011506:Proteins; D034741:RNA, Small Interfering; C011083:polyproline; D007791:Lactoylglutathione Lyase; D007213:Indomethacin