Mass Spectrometric Screening of Ligands with Lower Off-Rate from a Clicked-Based Pooled Library

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• 作者：Satoshi Arai ; Shota Hirosawa ; Yusuke Oguchi ; Madoka Suzuki ; Atsushi Murata ; Shin鈥檌chi Ishiwata ; Shinji Takeoka
• 刊名：ACS Combinatorial Science
• 出版年：2012
• 出版时间：August 13, 2012
• 年：2012
• 卷：14
• 期：8
• 页码：451-455
• 全文大小：289K
• 年卷期：v.14,no.8(August 13, 2012)
• ISSN：2156-8944

文摘

This paper describes a convenient screening method using ion trap electrospray ionization mass spectrometry to classify ligands to a target molecule in terms of kinetic parameters. We demonstrate this method in the screening of ligands to a hexahistidine tag from a pooled library synthesized by click chemistry. The ion trap mass spectrometry analysis revealed that higher stabilities of ligand-target complexes in the gas phase were related to lower dissociation rate constants, i.e., off-rates in solution. Finally, we prepared a fluorescent probe utilizing the ligand with lowest off-rate and succeeded in performing single molecule observations of hexahistidine-tagged myosin V walking on actin filaments.

Keywords:

click chemistry; fluorescent probes; high-throughput screening; mass spectrometry