LILBID laser dissociation curves: a mass spectrometry-based method for the quantitative assessment of dsDNA binding affinities

Abstract One current goal in native mass spectrometry is the assignment of binding affinities to noncovalent complexes. Here we introduce a novel implementation of the existing laser-induced liquid bead ion desorption (LILBID) mass spectrometry method: this new method, LILBID laser dissociation curv...

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Autores principales:	Phoebe Young, Genia Hense, Carina Immer, Jens Wöhnert, Nina Morgner
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/c7f5ac5f30ad48ce9d0d0f17b713962c
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spelling	oai:doaj.org-article:c7f5ac5f30ad48ce9d0d0f17b713962c2021-12-02T12:33:45ZLILBID laser dissociation curves: a mass spectrometry-based method for the quantitative assessment of dsDNA binding affinities10.1038/s41598-020-76867-92045-2322https://doaj.org/article/c7f5ac5f30ad48ce9d0d0f17b713962c2020-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-76867-9https://doaj.org/toc/2045-2322Abstract One current goal in native mass spectrometry is the assignment of binding affinities to noncovalent complexes. Here we introduce a novel implementation of the existing laser-induced liquid bead ion desorption (LILBID) mass spectrometry method: this new method, LILBID laser dissociation curves, assesses binding strengths quantitatively. In all LILBID applications, aqueous sample droplets are irradiated by 3 µm laser pulses. Variation of the laser energy transferred to the droplet during desorption affects the degree of complex dissociation. In LILBID laser dissociation curves, laser energy transfer is purposely varied, and a binding affinity is calculated from the resulting complex dissociation. A series of dsDNAs with different binding affinities was assessed using LILBID laser dissociation curves. The binding affinity results from the LILBID laser dissociation curves strongly correlated with the melting temperatures from UV melting curves and with dissociation constants from isothermal titration calorimetry, standard solution phase methods. LILBID laser dissociation curve data also showed good reproducibility and successfully predicted the melting temperatures and dissociation constants of three DNA sequences. LILBID laser dissociation curves are a promising native mass spectrometry binding affinity method, with reduced time and sample consumption compared to melting curves or titrations.Phoebe YoungGenia HenseCarina ImmerJens WöhnertNina MorgnerNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-13 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Phoebe Young Genia Hense Carina Immer Jens Wöhnert Nina Morgner LILBID laser dissociation curves: a mass spectrometry-based method for the quantitative assessment of dsDNA binding affinities
description	Abstract One current goal in native mass spectrometry is the assignment of binding affinities to noncovalent complexes. Here we introduce a novel implementation of the existing laser-induced liquid bead ion desorption (LILBID) mass spectrometry method: this new method, LILBID laser dissociation curves, assesses binding strengths quantitatively. In all LILBID applications, aqueous sample droplets are irradiated by 3 µm laser pulses. Variation of the laser energy transferred to the droplet during desorption affects the degree of complex dissociation. In LILBID laser dissociation curves, laser energy transfer is purposely varied, and a binding affinity is calculated from the resulting complex dissociation. A series of dsDNAs with different binding affinities was assessed using LILBID laser dissociation curves. The binding affinity results from the LILBID laser dissociation curves strongly correlated with the melting temperatures from UV melting curves and with dissociation constants from isothermal titration calorimetry, standard solution phase methods. LILBID laser dissociation curve data also showed good reproducibility and successfully predicted the melting temperatures and dissociation constants of three DNA sequences. LILBID laser dissociation curves are a promising native mass spectrometry binding affinity method, with reduced time and sample consumption compared to melting curves or titrations.
format	article
author	Phoebe Young Genia Hense Carina Immer Jens Wöhnert Nina Morgner
author_facet	Phoebe Young Genia Hense Carina Immer Jens Wöhnert Nina Morgner
author_sort	Phoebe Young
title	LILBID laser dissociation curves: a mass spectrometry-based method for the quantitative assessment of dsDNA binding affinities
title_short	LILBID laser dissociation curves: a mass spectrometry-based method for the quantitative assessment of dsDNA binding affinities
title_full	LILBID laser dissociation curves: a mass spectrometry-based method for the quantitative assessment of dsDNA binding affinities
title_fullStr	LILBID laser dissociation curves: a mass spectrometry-based method for the quantitative assessment of dsDNA binding affinities
title_full_unstemmed	LILBID laser dissociation curves: a mass spectrometry-based method for the quantitative assessment of dsDNA binding affinities
title_sort	lilbid laser dissociation curves: a mass spectrometry-based method for the quantitative assessment of dsdna binding affinities
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/c7f5ac5f30ad48ce9d0d0f17b713962c
work_keys_str_mv	AT phoebeyoung lilbidlaserdissociationcurvesamassspectrometrybasedmethodforthequantitativeassessmentofdsdnabindingaffinities AT geniahense lilbidlaserdissociationcurvesamassspectrometrybasedmethodforthequantitativeassessmentofdsdnabindingaffinities AT carinaimmer lilbidlaserdissociationcurvesamassspectrometrybasedmethodforthequantitativeassessmentofdsdnabindingaffinities AT jenswohnert lilbidlaserdissociationcurvesamassspectrometrybasedmethodforthequantitativeassessmentofdsdnabindingaffinities AT ninamorgner lilbidlaserdissociationcurvesamassspectrometrybasedmethodforthequantitativeassessmentofdsdnabindingaffinities
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LILBID laser dissociation curves: a mass spectrometry-based method for the quantitative assessment of dsDNA binding affinities

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