Mass spectrometric screening of ligands with lower off-rate from a clicked-based pooled library

Satoshi Arai, Shota Hirosawa, Yusuke Oguchi, Madoka Suzuki, Atsushi Murata, Shin'Ichi Ishiwata, Shinji Takeoka

Research output: Contribution to journalArticle

2 Citations (Scopus)

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 languageEnglish
Pages (from-to)451-455
Number of pages5
JournalACS Combinatorial Science
Volume14
Issue number8
DOIs
Publication statusPublished - 2012 Aug 13

Fingerprint

His-His-His-His-His-His
Screening
Ligands
Mass spectrometry
Myosin Type V
Ions
Electrospray ionization
Molecules
Fluorescent Dyes
Kinetic parameters
Actins
Rate constants
Gases

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 journalArticle

Arai, Satoshi ; Hirosawa, Shota ; Oguchi, Yusuke ; Suzuki, Madoka ; Murata, Atsushi ; Ishiwata, Shin'Ichi ; Takeoka, Shinji. / Mass spectrometric screening of ligands with lower off-rate from a clicked-based pooled library. In: ACS Combinatorial Science. 2012 ; Vol. 14, No. 8. pp. 451-455.
@article{bb0f9c2891214d0c99c1bb8dfc211061,
title = "Mass spectrometric screening of ligands with lower off-rate from a clicked-based pooled library",
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.",
keywords = "click chemistry, fluorescent probes, high-throughput screening, mass spectrometry",
author = "Satoshi Arai and Shota Hirosawa and Yusuke Oguchi and Madoka Suzuki and Atsushi Murata and Shin'Ichi Ishiwata and Shinji Takeoka",
year = "2012",
month = "8",
day = "13",
doi = "10.1021/co300028n",
language = "English",
volume = "14",
pages = "451--455",
journal = "ACS Combinatorial Science",
issn = "2156-8952",
publisher = "American Chemical Society",
number = "8",

}

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 -