Abstract
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.
Original language | English |
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Pages (from-to) | 451-455 |
Number of pages | 5 |
Journal | ACS Combinatorial Science |
Volume | 14 |
Issue number | 8 |
DOIs | |
Publication status | Published - 2012 Aug 13 |
Keywords
- click chemistry
- fluorescent probes
- high-throughput screening
- mass spectrometry
ASJC Scopus subject areas
- Chemistry(all)