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Unnatural amino acids increase activity and specificity of synthetic substrates for human and malarial cathepsin C
Mammalian cathepsin C is primarily responsible for the removal of N-terminal dipeptides and activation of several serine proteases in inflammatory or immune cells, while its malarial parasite ortholog dipeptidyl aminopeptidase 1 plays a crucial role in catabolizing the hemoglobin of its host erythro...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Vienna
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3962583/ https://www.ncbi.nlm.nih.gov/pubmed/24381006 http://dx.doi.org/10.1007/s00726-013-1654-2 |
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author | Poreba, Marcin Mihelic, Marko Krai, Priscilla Rajkovic, Jelena Krezel, Artur Pawelczak, Malgorzata Klemba, Michael Turk, Dusan Turk, Boris Latajka, Rafal Drag, Marcin |
author_facet | Poreba, Marcin Mihelic, Marko Krai, Priscilla Rajkovic, Jelena Krezel, Artur Pawelczak, Malgorzata Klemba, Michael Turk, Dusan Turk, Boris Latajka, Rafal Drag, Marcin |
author_sort | Poreba, Marcin |
collection | PubMed |
description | Mammalian cathepsin C is primarily responsible for the removal of N-terminal dipeptides and activation of several serine proteases in inflammatory or immune cells, while its malarial parasite ortholog dipeptidyl aminopeptidase 1 plays a crucial role in catabolizing the hemoglobin of its host erythrocyte. In this report, we describe the systematic substrate specificity analysis of three cathepsin C orthologs from Homo sapiens (human), Bos taurus (bovine) and Plasmodium falciparum (malaria parasite). Here, we present a new approach with a tailored fluorogenic substrate library designed and synthesized to probe the S1 and S2 pocket preferences of these enzymes with both natural and a broad range of unnatural amino acids. Our approach identified very efficiently hydrolyzed substrates containing unnatural amino acids, which resulted in the design of significantly better substrates than those previously known. Additionally, in this study significant differences in terms of the structures of optimal substrates for human and malarial orthologs are important from the therapeutic point of view. These data can be also used for the design of specific inhibitors or activity-based probes. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00726-013-1654-2) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-3962583 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Springer Vienna |
record_format | MEDLINE/PubMed |
spelling | pubmed-39625832014-03-24 Unnatural amino acids increase activity and specificity of synthetic substrates for human and malarial cathepsin C Poreba, Marcin Mihelic, Marko Krai, Priscilla Rajkovic, Jelena Krezel, Artur Pawelczak, Malgorzata Klemba, Michael Turk, Dusan Turk, Boris Latajka, Rafal Drag, Marcin Amino Acids Original Article Mammalian cathepsin C is primarily responsible for the removal of N-terminal dipeptides and activation of several serine proteases in inflammatory or immune cells, while its malarial parasite ortholog dipeptidyl aminopeptidase 1 plays a crucial role in catabolizing the hemoglobin of its host erythrocyte. In this report, we describe the systematic substrate specificity analysis of three cathepsin C orthologs from Homo sapiens (human), Bos taurus (bovine) and Plasmodium falciparum (malaria parasite). Here, we present a new approach with a tailored fluorogenic substrate library designed and synthesized to probe the S1 and S2 pocket preferences of these enzymes with both natural and a broad range of unnatural amino acids. Our approach identified very efficiently hydrolyzed substrates containing unnatural amino acids, which resulted in the design of significantly better substrates than those previously known. Additionally, in this study significant differences in terms of the structures of optimal substrates for human and malarial orthologs are important from the therapeutic point of view. These data can be also used for the design of specific inhibitors or activity-based probes. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00726-013-1654-2) contains supplementary material, which is available to authorized users. Springer Vienna 2014-01-01 2014 /pmc/articles/PMC3962583/ /pubmed/24381006 http://dx.doi.org/10.1007/s00726-013-1654-2 Text en © The Author(s) 2013 https://creativecommons.org/licenses/by/2.0/ Open AccessThis article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited. |
spellingShingle | Original Article Poreba, Marcin Mihelic, Marko Krai, Priscilla Rajkovic, Jelena Krezel, Artur Pawelczak, Malgorzata Klemba, Michael Turk, Dusan Turk, Boris Latajka, Rafal Drag, Marcin Unnatural amino acids increase activity and specificity of synthetic substrates for human and malarial cathepsin C |
title | Unnatural amino acids increase activity and specificity of synthetic substrates for human and malarial cathepsin C |
title_full | Unnatural amino acids increase activity and specificity of synthetic substrates for human and malarial cathepsin C |
title_fullStr | Unnatural amino acids increase activity and specificity of synthetic substrates for human and malarial cathepsin C |
title_full_unstemmed | Unnatural amino acids increase activity and specificity of synthetic substrates for human and malarial cathepsin C |
title_short | Unnatural amino acids increase activity and specificity of synthetic substrates for human and malarial cathepsin C |
title_sort | unnatural amino acids increase activity and specificity of synthetic substrates for human and malarial cathepsin c |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3962583/ https://www.ncbi.nlm.nih.gov/pubmed/24381006 http://dx.doi.org/10.1007/s00726-013-1654-2 |
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