Polyproline-Rod Approach to Isolating Protein Targets of Bioactive Small Molecules: Isolation of a New Target of Indomethacin

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• 作者：Shin-ichi Sato ; Youngjoo Kwon ; Shinji Kamisuki ; Neeta Srivastava ; Qian Mao ; Yoshinori Kawazoe ; Motonari Uesugi
• 刊名：Journal of the American Chemical Society
• 出版年：2007
• 出版时间：January 31, 2007
• 年：2007
• 卷：129
• 期：4
• 页码：873 - 880
• 全文大小：392K
• 年卷期：v.129,no.4(January 31, 2007)
• ISSN：1520-5126

文摘

Identification of protein targets of bioactive small molecules has been a technical hurdle ofchemical genetics. Here we report a polyproline-rod approach to isolating protein targets of small moleculesfrom 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 oflow-abundance or low-affinity proteins. In the course of the proof-of-concept experiments, we isolatedglyoxalase 1 (GLO1) as a new target of indomethacin, a widely used antiinflammatory drug. Molecularbiological experiments suggest that inhibition of GLO1 enzyme activity is related to the clinically recognizedbeneficial side effects of the indomethacin family of nonsteroidal antiinflammatory drugs.