Using Electrospray Ionization FTICR Mass Spectrometry To Study Competitive Binding of Inhibitors to Carbonic Anhydrase

Cheng, Xueheng; Chen, Ruidan; Bruce, James E.; Schwartz, Brenda L.; Anderson, Gordon A.; Hofstadler, Steven A.; Gale, David C.; Smith, Richard D.; Gao, Jinming; Sigal, George B.; Mammen, Mathai; Whitesides, George M.

doi:10.1021/ja00139a023

Cited by 187 publications

(148 citation statements)

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“…These features enable the determination of both the macroscopic and microscopic K a values for sequential binding of L to P. As a result, ESI-MS is ideally suited for characterizing allosteric binding. The ESI-MS assay also naturally lends itself to monitoring and quantifying protein-ligand interactions in solutions containing mixtures of ligands and/or proteins [25][26][27][28][29][30][31][32][33]. Not surprisingly, an emerging ESI-MS application is screening libraries of compounds against target proteins to identify specific interactions.…”

Section: Introductionmentioning

confidence: 99%

Reliable Determinations of Protein–Ligand Interactions by Direct ESI-MS Measurements. Are We There Yet?

Kitova

El-Hawiet

Schnier

et al. 2012

J. Am. Soc. Mass Spectrom.

222

321

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The association-dissociation of noncovalent interactions between protein and ligands, such as other proteins, carbohydrates, lipids, DNA, or small molecules, are critical events in many biological processes. The discovery and characterization of these interactions is essential to a complete understanding of biochemical reactions and pathways and to the design of novel therapeutic agents that may be used to treat a variety of diseases and infections. Over the last 20 y, electrospray ionization mass spectrometry (ESI-MS) has emerged as a versatile tool for the identification and quantification of protein-ligand interactions in vitro. Here, we describe the implementation of the direct ESI-MS assay for the determination of protein-ligand binding stoichiometry and affinity. Additionally, we outline common sources of error encountered with these measurements and various strategies to overcome them. Finally, we comment on some of the outstanding challenges associated with the implementation of the assay and highlight new areas where direct ESI-MS measurements are expected to make significant contributions in the future.

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Section: Introductionmentioning

confidence: 99%

Reliable Determinations of Protein–Ligand Interactions by Direct ESI-MS Measurements. Are We There Yet?

Kitova

El-Hawiet

Schnier

et al. 2012

J. Am. Soc. Mass Spectrom.

222

321

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“…The ability to measure multiple binding equilibria simultaneously makes the ESI-MS assay well suited for screening libraries of compounds against target proteins [38,39]. Where direct detection of the protein-ligand complexes by ESI-MS is not possible due to factors such as high molecular weight (MW) or protein heterogeneity, the catch-and-release (CaR) ESI-MS assay can be employed [40][41][42][43][44][45][46][47]. In this assay, ligand binding is established by releasing (as ions) ligands bound to the protein in the gas phase using activation methods such as collisioninduced dissociation (CID).…”

Section: Introductionmentioning

confidence: 99%

“…Typically, ligands can be identified based on their measured MW; in some cases, fragmentation of the released corresponding ligand ion or ion mobility separation (IMS) may be required for positive identification. The CaR-ESI-MS assay has been used to screen carbohydrate, peptide, and small molecule libraries against target proteins [40][41][42][43][44][45][46][47]. However, the reliability of the method has only been rigorously tested in the case of carbohydrate libraries [40,41,[45][46][47].…”

Section: Introductionmentioning

confidence: 99%

Screening Anti-Cancer Drugs against Tubulin using Catch-and-Release Electrospray Ionization Mass Spectrometry

Darestani

Winter

Kitova

et al. 2016

J. Am. Soc. Mass Spectrom.

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Abstract. Tubulin, which is the building block of microtubules, plays an important role in cell division. This critical role makes tubulin an attractive target for the development of chemotherapeutic drugs to treat cancer. Currently, there is no general binding assay for tubulin-drug interactions. The present work describes the application of the catch-and-release electrospray ionization mass spectrometry (CaR-ESI-MS) assay to investigate the binding of colchicinoid drugs to αβ-tubulin dimers extracted from porcine brain. Proof-of-concept experiments using positive (ligands with known affinities) and negative (non-binders) controls were performed to establish the reliability of the assay. The assay was then used to screen a library of seven colchicinoid analogues to test their binding to tubulin and to rank their affinities.

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“…In this application, a combinatorial library is pre-incubated with a receptor in solution and then analyzed directly using electrospray in order to identify receptor-ligand complexes in the gas phase [52][53][54][55][56]. Once a receptor-ligand complex is ionized and trapped in the FT-ICR mass spectrometer, the mass difference between the complex and the receptor alone might be measured with sufficient resolution and accuracy to determine the mass(es) and perhaps elemental composition(s) of the ligand(s).…”

Section: Screening Using Electrospray Fourier Transformion Cyclotron mentioning

confidence: 99%

Analysis and screening of combinatorial libraries using mass spectrometry

Shin

Breemen

2001

Biopharm & Drug Disp

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Mass spectrometry is a highly selective and high throughput analytical technique that is ideally suited for the identification and purity determination of large numbers of compounds prepared using combinatorial chemistry or for the dereplication of natural products. Compounds may be characterized based on molecular weight, elemental composition and structural features based on fragmentation patterns. When coupled to a separation technique such as highperformance liquid chromatography (HPLC) or capillary electrophoresis, mass spectrometric applications may be expanded to include analysis of complex mixtures. However, the slower speed of the separation step reduces the throughput of the analysis. This review concerns the application of mass spectrometry to the characterization of combinatorial libraries and the screening of library and natural product mixtures. Strategies to enhance the throughput of LC-MS are discussed including fast HPLC and parallel LC-MS. Also, mass spectrometry-based screening methods are described including frontal affinity chromatography-mass spectrometry, gel permeation chromatography LC-MS, direct electrospray mass spectrometry of receptor-ligand complexes, affinity chromatography-mass spectrometry, and pulsed ultrafiltration mass spectrometry.

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Using Electrospray Ionization FTICR Mass Spectrometry To Study Competitive Binding of Inhibitors to Carbonic Anhydrase

Cited by 187 publications

References 0 publications

Reliable Determinations of Protein–Ligand Interactions by Direct ESI-MS Measurements. Are We There Yet?

Reliable Determinations of Protein–Ligand Interactions by Direct ESI-MS Measurements. Are We There Yet?

Screening Anti-Cancer Drugs against Tubulin using Catch-and-Release Electrospray Ionization Mass Spectrometry

Analysis and screening of combinatorial libraries using mass spectrometry

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