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A novel and highly efficient purification procedure for native human dipeptidyl peptidase 3 from human blood cell lysate
Dipeptidyl amino-peptidase 3 (DPP3) is an aminopeptidase involved in peptide degradation, including hormone peptides as angiotensin II and enkephalins. DPP3 plasma activity increases in septic patients and correlates with mortality risk. However, the exact physiological role of DPP3 remains unclear...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6685676/ https://www.ncbi.nlm.nih.gov/pubmed/31390378 http://dx.doi.org/10.1371/journal.pone.0220866 |
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author | Kaufmann, Paul Muenzner, Matthias Kästorf, Mandy Santos, Karine Hartmann, Tobias Dienelt, Anke Rehfeld, Linda Bergmann, Andreas |
author_facet | Kaufmann, Paul Muenzner, Matthias Kästorf, Mandy Santos, Karine Hartmann, Tobias Dienelt, Anke Rehfeld, Linda Bergmann, Andreas |
author_sort | Kaufmann, Paul |
collection | PubMed |
description | Dipeptidyl amino-peptidase 3 (DPP3) is an aminopeptidase involved in peptide degradation, including hormone peptides as angiotensin II and enkephalins. DPP3 plasma activity increases in septic patients and correlates with mortality risk. However, the exact physiological role of DPP3 remains unclear and animal studies are necessary to reveal the function of DPP3 in vivo. To this demand, we developed a two-step purification procedure for isolation of native human DPP3 from blood cell lysate (BCL) that is suitable for in vivo applications. With the use of monoclonal antibodies coupled to beads in combination with an ion-exchange chromatography, we recovered 68% of human DPP3 activity from BCL with a purity of ≥ 95%. Purified human DPP3 was assayed for activity and protein concentration using recently published DPP3-activity- and immunoassays. Additionally, protein stability and storage in relevant buffers were tested. Our results provide a promising strategy for fast and efficient isolation of human DPP3. The purified human DPP3 represents the native state of DPP3, suitable for future in vivo applications to investigate the physiological role of DPP3 and its involvement in pathophysiological conditions. |
format | Online Article Text |
id | pubmed-6685676 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-66856762019-08-15 A novel and highly efficient purification procedure for native human dipeptidyl peptidase 3 from human blood cell lysate Kaufmann, Paul Muenzner, Matthias Kästorf, Mandy Santos, Karine Hartmann, Tobias Dienelt, Anke Rehfeld, Linda Bergmann, Andreas PLoS One Research Article Dipeptidyl amino-peptidase 3 (DPP3) is an aminopeptidase involved in peptide degradation, including hormone peptides as angiotensin II and enkephalins. DPP3 plasma activity increases in septic patients and correlates with mortality risk. However, the exact physiological role of DPP3 remains unclear and animal studies are necessary to reveal the function of DPP3 in vivo. To this demand, we developed a two-step purification procedure for isolation of native human DPP3 from blood cell lysate (BCL) that is suitable for in vivo applications. With the use of monoclonal antibodies coupled to beads in combination with an ion-exchange chromatography, we recovered 68% of human DPP3 activity from BCL with a purity of ≥ 95%. Purified human DPP3 was assayed for activity and protein concentration using recently published DPP3-activity- and immunoassays. Additionally, protein stability and storage in relevant buffers were tested. Our results provide a promising strategy for fast and efficient isolation of human DPP3. The purified human DPP3 represents the native state of DPP3, suitable for future in vivo applications to investigate the physiological role of DPP3 and its involvement in pathophysiological conditions. Public Library of Science 2019-08-07 /pmc/articles/PMC6685676/ /pubmed/31390378 http://dx.doi.org/10.1371/journal.pone.0220866 Text en © 2019 Kaufmann et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Kaufmann, Paul Muenzner, Matthias Kästorf, Mandy Santos, Karine Hartmann, Tobias Dienelt, Anke Rehfeld, Linda Bergmann, Andreas A novel and highly efficient purification procedure for native human dipeptidyl peptidase 3 from human blood cell lysate |
title | A novel and highly efficient purification procedure for native human dipeptidyl peptidase 3 from human blood cell lysate |
title_full | A novel and highly efficient purification procedure for native human dipeptidyl peptidase 3 from human blood cell lysate |
title_fullStr | A novel and highly efficient purification procedure for native human dipeptidyl peptidase 3 from human blood cell lysate |
title_full_unstemmed | A novel and highly efficient purification procedure for native human dipeptidyl peptidase 3 from human blood cell lysate |
title_short | A novel and highly efficient purification procedure for native human dipeptidyl peptidase 3 from human blood cell lysate |
title_sort | novel and highly efficient purification procedure for native human dipeptidyl peptidase 3 from human blood cell lysate |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6685676/ https://www.ncbi.nlm.nih.gov/pubmed/31390378 http://dx.doi.org/10.1371/journal.pone.0220866 |
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