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Fenton-Chemistry-Based Oxidative Modification of Proteins Reflects Their Conformation

Nehls, Thomas ; Heymann, Tim ; Meyners, Christian ; Hausch, Felix ; Lermyte, Frederik (2022)
Fenton-Chemistry-Based Oxidative Modification of Proteins Reflects Their Conformation.
In: International Journal of Molecular Sciences, 2022, 22 (18)
doi: 10.26083/tuprints-00021257
Article, Secondary publication, Publisher's Version

Abstract

In order to understand protein structure to a sufficient extent for, e.g., drug discovery, no single technique can provide satisfactory information on both the lowest-energy conformation and on dynamic changes over time (the ‘four-dimensional’ protein structure). Instead, a combination of complementary techniques is required. Mass spectrometry methods have shown promise in addressing protein dynamics, but often rely on the use of high-end commercial or custom instruments. Here, we apply well-established chemistry to conformation-sensitive oxidative protein labelling on a timescale of a few seconds, followed by analysis through a routine protein analysis workflow. For a set of model proteins, we show that site selectivity of labelling can indeed be rationalised in terms of known structural information, and that conformational changes induced by ligand binding are reflected in the modification pattern. In addition to conventional bottom-up analysis, further insights are obtained from intact mass measurement and native mass spectrometry. We believe that this method will provide a valuable and robust addition to the ‘toolbox’ of mass spectrometry researchers studying higher-order protein structure.

Item Type: Article
Erschienen: 2022
Creators: Nehls, Thomas ; Heymann, Tim ; Meyners, Christian ; Hausch, Felix ; Lermyte, Frederik
Type of entry: Secondary publication
Title: Fenton-Chemistry-Based Oxidative Modification of Proteins Reflects Their Conformation
Language: English
Date: 2022
Year of primary publication: 2022
Publisher: MDPI
Journal or Publication Title: International Journal of Molecular Sciences
Volume of the journal: 22
Issue Number: 18
Collation: 18 Seiten
DOI: 10.26083/tuprints-00021257
URL / URN: https://tuprints.ulb.tu-darmstadt.de/21257
Corresponding Links:
Origin: Secondary publication via sponsored Golden Open Access
Abstract:

In order to understand protein structure to a sufficient extent for, e.g., drug discovery, no single technique can provide satisfactory information on both the lowest-energy conformation and on dynamic changes over time (the ‘four-dimensional’ protein structure). Instead, a combination of complementary techniques is required. Mass spectrometry methods have shown promise in addressing protein dynamics, but often rely on the use of high-end commercial or custom instruments. Here, we apply well-established chemistry to conformation-sensitive oxidative protein labelling on a timescale of a few seconds, followed by analysis through a routine protein analysis workflow. For a set of model proteins, we show that site selectivity of labelling can indeed be rationalised in terms of known structural information, and that conformational changes induced by ligand binding are reflected in the modification pattern. In addition to conventional bottom-up analysis, further insights are obtained from intact mass measurement and native mass spectrometry. We believe that this method will provide a valuable and robust addition to the ‘toolbox’ of mass spectrometry researchers studying higher-order protein structure.

Status: Publisher's Version
URN: urn:nbn:de:tuda-tuprints-212576
Additional Information:

Data Availability Statement: The data and materials underlying this article will be shared on request to the corresponding author.

Keywords: mass spectrometry; protein folding; protein–ligand interactions; protein dynamics; FK506-binding protein; FKBP12; FKBP51

Classification DDC: 500 Science and mathematics > 540 Chemistry
500 Science and mathematics > 570 Life sciences, biology
Divisions: 07 Department of Chemistry
07 Department of Chemistry > Clemens-Schöpf-Institut > Fachgebiet Biochemie
Date Deposited: 06 May 2022 12:06
Last Modified: 09 May 2022 06:27
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