Analysis of Combinatorial Libraries Using Electrospray Fourier Transform Ion Cyclotron Resonance Mass Spectrometry

Nawrocki, Joseph P.; Wigger, Maria; Wilson, Clifford H.; Hayes, Thomas W.; Senko, Michael W.; Benner, Steven A.; Eyler, John R.

doi:10.1002/(sici)1097-0231(199611)10:14<1860::aid-rcm770>3.3.co;2-r

Cited by 21 publications

(37 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We recently reported the use of ESI FT-ICR mass spectrometry as a tool to analyze highly degenerate libraries of compounds [31], and showed how it can be applied to the real time monitoring of substrate library screening by enzymes [11]. However, many biological processes can be triggered simply by non-covalent binding of a ligand to a target biomolecule without any further covalent modification.…”

Section: Resultsmentioning

confidence: 99%

Fourier transform-ion cyclotron resonance mass spectrometric resolution, identification, and screening of non-covalent complexes of Hck Src homology 2 domain receptor and ligands from a 324-member peptide combinatorial library

Wigger

Eyler

Benner

et al. 2002

J. Am. Soc. Mass Spectrom.

Self Cite

View full text Add to dashboard Cite

The preferred ligands for the Hck Src homology 2 domain among a combinatorial library containing 324 different peptides were determined in a single experiment involving Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry (MS), electrospray ionization (ESI), stored-waveform inverse Fourier transformation (SWIFT), and infrared multiphoton laser disassociation (IRMPD). These were compared with the results obtained by conventional screening of the peptide library in solution using affinity chromatography. The results reported here show that by combining ESI, FT-ICR MS, SWIFT, and IRMPD, ligands likely to bind under physiological conditions are rapidly and efficiently identified, even from complex library mixtures. In the gas phase some discrimination against hydrophobic ligands could be observed. However, the illustrated feasibility of identifying high affinity ligand via gas-phase screening of complex library mixtures should lead to broad applications in the development of ligands for proteins with interesting biological activity, the first step that must be taken to develop a therapeutic agent. Important in the application of FT-ICR methods to biological macromolecules are "soft" ionization techniques, which generate macromolecular ions without fragmentation. Electrospray ionization (ESI) and matrix-assisted laser desorption/ionization (MALDI) are two of these [4,5]. ESI involves spraying an aqueous solution of a biological macromolecule under "quasiphysiological" conditions directly into a mass spectrometer. The solvent evaporates in the vacuum from the droplets, gently "elevating" a biological ion from its native environment in solution. This process frequently leaves non-covalent macromolecular complexes intact and allows the mass spectrometric observation of enzyme-substrate, receptor-ligand, peptide-peptide, protein-subunit, oligonucleotide-toxin/drug, and DNA duplex binding in the gas phase [6].In many cases, binding constants estimated from mass spectrometric studies match the binding constants obtained in solution to within an order of magnitude [7]. The gentleness of ESI allows H/D exchange experiments in the gas phase to give information about protein conformation and protein/protein interactions based on measurements of the exchange rates of protein backbone amide protons [8,9]. This combination of

show abstract

Section: Resultsmentioning

confidence: 99%

Fourier transform-ion cyclotron resonance mass spectrometric resolution, identification, and screening of non-covalent complexes of Hck Src homology 2 domain receptor and ligands from a 324-member peptide combinatorial library

Wigger

Eyler

Benner

et al. 2002

J. Am. Soc. Mass Spectrom.

Self Cite

View full text Add to dashboard Cite

show abstract

“…The absolute mean errors were 5.2 (σ = 7.4 ppm), 0.7 (σ = 0.9 ppm), and 0.8 (σ = 1.0 ppm) ppm for the external, conventional internal, and dual ionization internal calibrations, respectively. In another application, Nawrocki et al [82] employed a 4.7-T external-source ESI FT-ICR mass spectrometer to analyze small-peptide libraries, demonstrating the feasibility of analyzing several combinatorial libraries containing 100 to 10,000 small peptides. Furthermore, by comparing the FTMS data with computer-simulated combinatorial library mass spectra, the authors were able to monitor the diversity and degeneracy of the library syntheses.…”

Section: High-throughput Lc/ms For Combinatorial Chemistrymentioning

confidence: 99%

LC/MS: Theory, Instrumentation, and Applications to Small Molecules

Chen¹,

Zhang²,

Pramanik³

2006

HPLC for Pharmaceutical Scientists

View full text Add to dashboard Cite

“…Two of the earliest reports of the successful application of FTICR-MS to combinatorial libraries were published by Winger and Campana of the Finnigan FTMS group in 1996 [26] and by Brenner and coworkers of the University of Florida [27] at about the same time. Winger and Campana demonstrated the screening of the octapeptide mixture SIIN-X-EKL (where X can represents any of the 19 amino acids, except cysteine) by use of positive-ion ESI FTICR-MS, without the need for chromatographic separation [26].…”

Section: Direct Fticr-ms Analysis Of Combinatorial Librariesmentioning

confidence: 99%

“…Benner and coworkers [27] demonstrated the utility of FTICR-MS for analysis of selected small-peptide combinatorial libraries containing 10 2 to 10 4 individual components. They also simulated the synthesized libraries by using a computer program which enabled information to be extracted about the degeneracy and diversity of the libraries.…”

Section: Direct Fticr-ms Analysis Of Combinatorial Librariesmentioning

confidence: 99%

Ultra-high resolution for mass spectrometric analysis of complex and low-abundance mixtures – the emergence of FTICR-MS as an essential analytical tool

2002

View full text Add to dashboard Cite

The role of high resolution in mass spectrometric analysisThe development of applications of mass spectrometry to chemical analysis has followed several paths, such as invention of new techniques for ionization, for acquisition of additional information concerning the relationship between fragment ions in the spectrum (tandem mass spectrometry), and for on-line coupling of a mass spectrometer to separatory techniques for real-time mixture analysis. Other features of any analytical instrument, such as sensitivity and detection limits, spectral acquisition rate, m/z range, etc., have, in mass spectrometry, often been driven by the need to match the low sample levels and higher resolution which have resulted from the introduction of capillary-based chromatographic methods.This article, however, is concerned with another figure-of-merit, the resolving power of the mass spectrometer itself. Historically, the need for higher resolving power was driven by investigations into isotopic compositions of the elements after Aston's landmark discoveries. Magnetic sector instruments were the dominant technology, and the invention of the double-focusing principle enabled simultaneous achievement of large increases in sensitivity and resolving power, counter-intuitive for a dispersion analyzer. The desire to detect and measure extremely low levels of rare isotopes, the mass spectral peaks of which were adjacent to those of much more abundant isotopes, required high resolving power, defined in terms of peak widths near the base of the peaks, to achieve the required "abundance sensitivity" for the rare

show abstract

Analysis of Combinatorial Libraries Using Electrospray Fourier Transform Ion Cyclotron Resonance Mass Spectrometry

Cited by 21 publications

References 0 publications

Fourier transform-ion cyclotron resonance mass spectrometric resolution, identification, and screening of non-covalent complexes of Hck Src homology 2 domain receptor and ligands from a 324-member peptide combinatorial library

Fourier transform-ion cyclotron resonance mass spectrometric resolution, identification, and screening of non-covalent complexes of Hck Src homology 2 domain receptor and ligands from a 324-member peptide combinatorial library

LC/MS: Theory, Instrumentation, and Applications to Small Molecules

Ultra-high resolution for mass spectrometric analysis of complex and low-abundance mixtures – the emergence of FTICR-MS as an essential analytical tool

Contact Info

Product

Resources

About