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Compactness of Protein Folds Alters Disulfide‐Bond Reducibility by Three Orders of Magnitude: A Comprehensive Kinetic Case Study on the Reduction of Differently Sized Tryptophan Cage Model Proteins

A new approach to monitor disulfide‐bond reduction in the vicinity of aromatic cluster(s) has been derived by using the near‐UV range (λ=266–293 nm) of electronic circular dichroism (ECD) spectra. By combining the results from NMR and ECD spectroscopy, the 3D fold characteristics and associated redu...

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Autores principales: Horváth, Dániel, Taricska, Nóra, Keszei, Ernő, Stráner, Pál, Farkas, Viktor, Tóth, Gábor K., Perczel, András
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7079008/
https://www.ncbi.nlm.nih.gov/pubmed/31475422
http://dx.doi.org/10.1002/cbic.201900470
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author Horváth, Dániel
Taricska, Nóra
Keszei, Ernő
Stráner, Pál
Farkas, Viktor
Tóth, Gábor K.
Perczel, András
author_facet Horváth, Dániel
Taricska, Nóra
Keszei, Ernő
Stráner, Pál
Farkas, Viktor
Tóth, Gábor K.
Perczel, András
author_sort Horváth, Dániel
collection PubMed
description A new approach to monitor disulfide‐bond reduction in the vicinity of aromatic cluster(s) has been derived by using the near‐UV range (λ=266–293 nm) of electronic circular dichroism (ECD) spectra. By combining the results from NMR and ECD spectroscopy, the 3D fold characteristics and associated reduction rate constants (k) of E19_SS, which is a highly thermostable, disulfide‐bond reinforced 39‐amino acid long exenatide mimetic, and its N‐terminally truncated derivatives have been determined under different experimental conditions. Single disulfide bond reduction of the E19_SS model (with an 18‐fold excess of tris(2‐carboxyethyl)phosphine, pH 7, 37 °C) takes hours, which is 20–30 times longer than that expected, and thus, would not reach completion by applying commonly used reduction protocols. It is found that structural, steric, and electrostatic factors influence the reduction rate, resulting in orders of magnitude differences in reduction half‐lives (900>t (1/2)>1 min) even for structurally similar, well‐folded derivatives of a small model protein.
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spelling pubmed-70790082020-03-19 Compactness of Protein Folds Alters Disulfide‐Bond Reducibility by Three Orders of Magnitude: A Comprehensive Kinetic Case Study on the Reduction of Differently Sized Tryptophan Cage Model Proteins Horváth, Dániel Taricska, Nóra Keszei, Ernő Stráner, Pál Farkas, Viktor Tóth, Gábor K. Perczel, András Chembiochem Full Papers A new approach to monitor disulfide‐bond reduction in the vicinity of aromatic cluster(s) has been derived by using the near‐UV range (λ=266–293 nm) of electronic circular dichroism (ECD) spectra. By combining the results from NMR and ECD spectroscopy, the 3D fold characteristics and associated reduction rate constants (k) of E19_SS, which is a highly thermostable, disulfide‐bond reinforced 39‐amino acid long exenatide mimetic, and its N‐terminally truncated derivatives have been determined under different experimental conditions. Single disulfide bond reduction of the E19_SS model (with an 18‐fold excess of tris(2‐carboxyethyl)phosphine, pH 7, 37 °C) takes hours, which is 20–30 times longer than that expected, and thus, would not reach completion by applying commonly used reduction protocols. It is found that structural, steric, and electrostatic factors influence the reduction rate, resulting in orders of magnitude differences in reduction half‐lives (900>t (1/2)>1 min) even for structurally similar, well‐folded derivatives of a small model protein. John Wiley and Sons Inc. 2019-11-18 2020-03-02 /pmc/articles/PMC7079008/ /pubmed/31475422 http://dx.doi.org/10.1002/cbic.201900470 Text en © 2019 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Full Papers
Horváth, Dániel
Taricska, Nóra
Keszei, Ernő
Stráner, Pál
Farkas, Viktor
Tóth, Gábor K.
Perczel, András
Compactness of Protein Folds Alters Disulfide‐Bond Reducibility by Three Orders of Magnitude: A Comprehensive Kinetic Case Study on the Reduction of Differently Sized Tryptophan Cage Model Proteins
title Compactness of Protein Folds Alters Disulfide‐Bond Reducibility by Three Orders of Magnitude: A Comprehensive Kinetic Case Study on the Reduction of Differently Sized Tryptophan Cage Model Proteins
title_full Compactness of Protein Folds Alters Disulfide‐Bond Reducibility by Three Orders of Magnitude: A Comprehensive Kinetic Case Study on the Reduction of Differently Sized Tryptophan Cage Model Proteins
title_fullStr Compactness of Protein Folds Alters Disulfide‐Bond Reducibility by Three Orders of Magnitude: A Comprehensive Kinetic Case Study on the Reduction of Differently Sized Tryptophan Cage Model Proteins
title_full_unstemmed Compactness of Protein Folds Alters Disulfide‐Bond Reducibility by Three Orders of Magnitude: A Comprehensive Kinetic Case Study on the Reduction of Differently Sized Tryptophan Cage Model Proteins
title_short Compactness of Protein Folds Alters Disulfide‐Bond Reducibility by Three Orders of Magnitude: A Comprehensive Kinetic Case Study on the Reduction of Differently Sized Tryptophan Cage Model Proteins
title_sort compactness of protein folds alters disulfide‐bond reducibility by three orders of magnitude: a comprehensive kinetic case study on the reduction of differently sized tryptophan cage model proteins
topic Full Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7079008/
https://www.ncbi.nlm.nih.gov/pubmed/31475422
http://dx.doi.org/10.1002/cbic.201900470
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