Mass spectrometric screening of ligands with lower off-rate from a clicked-based pooled library.
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.
Satoshi
S
Arai
Shota
S
Hirosawa
Yusuke
Y
Oguchi
Madoka
M
Suzuki
Atsushi
A
Murata
Shin'ichi
S
Ishiwata
Shinji
S
Takeoka
D057930:Click Chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D008024:Ligands; D054852:Small Molecule Libraries / Q000138:chemical synthesis / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization
D005456:Fluorescent Dyes; D008024:Ligands; D054852:Small Molecule Libraries