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Predominant cleavage of proteins N-terminal to serines and threonines using scandium(III) triflate
ABSTRACT: Proteolytic digestion prior to LC–MS analysis is a key step for the identification of proteins. Digestion of proteins is typically performed with trypsin, but certain proteins or important protein sequence regions might be missed using this endoproteinase. Only few alternative endoproteina...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7064626/ https://www.ncbi.nlm.nih.gov/pubmed/31667593 http://dx.doi.org/10.1007/s00775-019-01733-7 |
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author | Koehler, Christian J. Thiede, Bernd |
author_facet | Koehler, Christian J. Thiede, Bernd |
author_sort | Koehler, Christian J. |
collection | PubMed |
description | ABSTRACT: Proteolytic digestion prior to LC–MS analysis is a key step for the identification of proteins. Digestion of proteins is typically performed with trypsin, but certain proteins or important protein sequence regions might be missed using this endoproteinase. Only few alternative endoproteinases are available and chemical cleavage of proteins is rarely used. Recently, it has been reported that some metal complexes can act as artificial proteases. In particular, the Lewis acid scandium(III) triflate has been shown to catalyze the cleavage of peptide bonds to serine and threonine residues. Therefore, we investigated if this compound can also be used for the cleavage of proteins. For this purpose, several single proteins, the 20S immune-proteasome (17 proteins), and the Universal Proteomics Standard UPS1 (48 proteins) were analyzed by MALDI–MS and/or LC–MS. A high cleavage specificity N-terminal to serine and threonine residues was observed, but also additional peptides with deviating cleavage specificity were found. Scandium(III) triflate can be a useful tool in protein analysis as no other reagent has been reported yet which showed cleavage specificity within proteins to serines and threonines. GRAPHIC ABSTRACT: [Image: see text] ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00775-019-01733-7) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-7064626 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-70646262020-03-23 Predominant cleavage of proteins N-terminal to serines and threonines using scandium(III) triflate Koehler, Christian J. Thiede, Bernd J Biol Inorg Chem Original Paper ABSTRACT: Proteolytic digestion prior to LC–MS analysis is a key step for the identification of proteins. Digestion of proteins is typically performed with trypsin, but certain proteins or important protein sequence regions might be missed using this endoproteinase. Only few alternative endoproteinases are available and chemical cleavage of proteins is rarely used. Recently, it has been reported that some metal complexes can act as artificial proteases. In particular, the Lewis acid scandium(III) triflate has been shown to catalyze the cleavage of peptide bonds to serine and threonine residues. Therefore, we investigated if this compound can also be used for the cleavage of proteins. For this purpose, several single proteins, the 20S immune-proteasome (17 proteins), and the Universal Proteomics Standard UPS1 (48 proteins) were analyzed by MALDI–MS and/or LC–MS. A high cleavage specificity N-terminal to serine and threonine residues was observed, but also additional peptides with deviating cleavage specificity were found. Scandium(III) triflate can be a useful tool in protein analysis as no other reagent has been reported yet which showed cleavage specificity within proteins to serines and threonines. GRAPHIC ABSTRACT: [Image: see text] ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00775-019-01733-7) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2019-10-30 2020 /pmc/articles/PMC7064626/ /pubmed/31667593 http://dx.doi.org/10.1007/s00775-019-01733-7 Text en © The Author(s) 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. |
spellingShingle | Original Paper Koehler, Christian J. Thiede, Bernd Predominant cleavage of proteins N-terminal to serines and threonines using scandium(III) triflate |
title | Predominant cleavage of proteins N-terminal to serines and threonines using scandium(III) triflate |
title_full | Predominant cleavage of proteins N-terminal to serines and threonines using scandium(III) triflate |
title_fullStr | Predominant cleavage of proteins N-terminal to serines and threonines using scandium(III) triflate |
title_full_unstemmed | Predominant cleavage of proteins N-terminal to serines and threonines using scandium(III) triflate |
title_short | Predominant cleavage of proteins N-terminal to serines and threonines using scandium(III) triflate |
title_sort | predominant cleavage of proteins n-terminal to serines and threonines using scandium(iii) triflate |
topic | Original Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7064626/ https://www.ncbi.nlm.nih.gov/pubmed/31667593 http://dx.doi.org/10.1007/s00775-019-01733-7 |
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