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Identification of Protease Specificity Using Biotin-Labeled Substrates
BACKGROUND: Proteolysis constitutes a major post-translational modification. For example, proteases regulate the activation or inactivation of various proteins, such as enzymes, growth factors, and peptide hormones. Proteases have substrate specificity, and protease expression regulates the specific...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Bentham Open
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5418938/ https://www.ncbi.nlm.nih.gov/pubmed/28567123 http://dx.doi.org/10.2174/1874091X01711010027 |
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author | Yamamoto, Hiroyuki Saito, Syota Sawaguchi, Yoshikazu Kimura, Michio |
author_facet | Yamamoto, Hiroyuki Saito, Syota Sawaguchi, Yoshikazu Kimura, Michio |
author_sort | Yamamoto, Hiroyuki |
collection | PubMed |
description | BACKGROUND: Proteolysis constitutes a major post-translational modification. For example, proteases regulate the activation or inactivation of various proteins, such as enzymes, growth factors, and peptide hormones. Proteases have substrate specificity, and protease expression regulates the specific and regional activation or inactivation of several functional proteins. METHODS: We demonstrate a novel method for determining protease specificity through the use of MALDI-TOF mass spectrometry with biotin-labeled substrates. RESULTS: This method was able to determine the specificity of TPCK-trypsin, V8 protease, elastase and cyanogen bromide cleavage, and the results were similar to previous reports. In addition, the method can be used to measure crude samples, such as tumor extracts. CONCLUSION: We demonstrated that this method could identify protease specificity after simple processing, even for crude samples. |
format | Online Article Text |
id | pubmed-5418938 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Bentham Open |
record_format | MEDLINE/PubMed |
spelling | pubmed-54189382017-05-31 Identification of Protease Specificity Using Biotin-Labeled Substrates Yamamoto, Hiroyuki Saito, Syota Sawaguchi, Yoshikazu Kimura, Michio Open Biochem J Article BACKGROUND: Proteolysis constitutes a major post-translational modification. For example, proteases regulate the activation or inactivation of various proteins, such as enzymes, growth factors, and peptide hormones. Proteases have substrate specificity, and protease expression regulates the specific and regional activation or inactivation of several functional proteins. METHODS: We demonstrate a novel method for determining protease specificity through the use of MALDI-TOF mass spectrometry with biotin-labeled substrates. RESULTS: This method was able to determine the specificity of TPCK-trypsin, V8 protease, elastase and cyanogen bromide cleavage, and the results were similar to previous reports. In addition, the method can be used to measure crude samples, such as tumor extracts. CONCLUSION: We demonstrated that this method could identify protease specificity after simple processing, even for crude samples. Bentham Open 2017-04-21 /pmc/articles/PMC5418938/ /pubmed/28567123 http://dx.doi.org/10.2174/1874091X01711010027 Text en © 2017 Yamamoto et al. https://creativecommons.org/licenses/by/4.0/legalcode This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: https://creativecommons.org/licenses/by/4.0/legalcode. This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Article Yamamoto, Hiroyuki Saito, Syota Sawaguchi, Yoshikazu Kimura, Michio Identification of Protease Specificity Using Biotin-Labeled Substrates |
title | Identification of Protease Specificity Using Biotin-Labeled Substrates |
title_full | Identification of Protease Specificity Using Biotin-Labeled Substrates |
title_fullStr | Identification of Protease Specificity Using Biotin-Labeled Substrates |
title_full_unstemmed | Identification of Protease Specificity Using Biotin-Labeled Substrates |
title_short | Identification of Protease Specificity Using Biotin-Labeled Substrates |
title_sort | identification of protease specificity using biotin-labeled substrates |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5418938/ https://www.ncbi.nlm.nih.gov/pubmed/28567123 http://dx.doi.org/10.2174/1874091X01711010027 |
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