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 |
|---|---|
| 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 |
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Keywords
- click chemistry
- fluorescent probes
- high-throughput screening
- mass spectrometry
ASJC Scopus subject areas
- Chemistry(all)
Cite this
Mass spectrometric screening of ligands with lower off-rate from a clicked-based pooled library. / Arai, Satoshi; Hirosawa, Shota; Oguchi, Yusuke; Suzuki, Madoka; Murata, Atsushi; Ishiwata, Shin'Ichi; Takeoka, Shinji.
In: ACS Combinatorial Science, Vol. 14, No. 8, 13.08.2012, p. 451-455.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Mass spectrometric screening of ligands with lower off-rate from a clicked-based pooled library
AU - Arai, Satoshi
AU - Hirosawa, Shota
AU - Oguchi, Yusuke
AU - Suzuki, Madoka
AU - Murata, Atsushi
AU - Ishiwata, Shin'Ichi
AU - Takeoka, Shinji
PY - 2012/8/13
Y1 - 2012/8/13
N2 - 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.
AB - 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.
KW - click chemistry
KW - fluorescent probes
KW - high-throughput screening
KW - mass spectrometry
UR - http://www.scopus.com/inward/record.url?scp=84864999758&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84864999758&partnerID=8YFLogxK
U2 - 10.1021/co300028n
DO - 10.1021/co300028n
M3 - Article
C2 - 22809242
AN - SCOPUS:84864999758
VL - 14
SP - 451
EP - 455
JO - ACS Combinatorial Science
JF - ACS Combinatorial Science
SN - 2156-8952
IS - 8
ER -