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MMP activation–associated aminopeptidase N reveals a bivalent 14-3-3 binding motif
Aminopeptidase N (APN, CD13) is a transmembrane ectopeptidase involved in many crucial cellular functions. Besides its role as a peptidase, APN also mediates signal transduction and is involved in the activation of matrix metalloproteinases (MMPs). MMPs function in tissue remodeling within the extra...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7939381/ https://www.ncbi.nlm.nih.gov/pubmed/33109610 http://dx.doi.org/10.1074/jbc.RA120.014708 |
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author | Kiehstaller, Sebastian Ottmann, Christian Hennig, Sven |
author_facet | Kiehstaller, Sebastian Ottmann, Christian Hennig, Sven |
author_sort | Kiehstaller, Sebastian |
collection | PubMed |
description | Aminopeptidase N (APN, CD13) is a transmembrane ectopeptidase involved in many crucial cellular functions. Besides its role as a peptidase, APN also mediates signal transduction and is involved in the activation of matrix metalloproteinases (MMPs). MMPs function in tissue remodeling within the extracellular space and are therefore involved in many human diseases, such as fibrosis, rheumatoid arthritis, tumor angiogenesis, and metastasis, as well as viral infections. However, the exact mechanism that leads to APN-driven MMP activation is unclear. It was previously shown that extracellular 14-3-3 adapter proteins bind to APN and thereby induce the transcription of MMPs. As a first step, we sought to identify potential 14-3-3–binding sites in the APN sequence. We constructed a set of phosphorylated peptides derived from APN to probe for interactions. We identified and characterized a canonical 14-3-3–binding site (site 1) within the flexible, structurally unresolved N-terminal APN region using direct binding fluorescence polarization assays and thermodynamic analysis. In addition, we identified a secondary, noncanonical binding site (site 2), which enhances the binding affinity in combination with site 1 by many orders of magnitude. Finally, we solved crystal structures of 14-3-3σ bound to mono- and bis-phosphorylated APN-derived peptides, which revealed atomic details of the binding mode of mono- and bivalent 14-3-3 interactions. Therefore, our findings shed some light on the first steps of APN-mediated MMP activation and open the field for further investigation of this important signaling pathway. |
format | Online Article Text |
id | pubmed-7939381 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-79393812021-06-08 MMP activation–associated aminopeptidase N reveals a bivalent 14-3-3 binding motif Kiehstaller, Sebastian Ottmann, Christian Hennig, Sven J Biol Chem Signal Transduction Aminopeptidase N (APN, CD13) is a transmembrane ectopeptidase involved in many crucial cellular functions. Besides its role as a peptidase, APN also mediates signal transduction and is involved in the activation of matrix metalloproteinases (MMPs). MMPs function in tissue remodeling within the extracellular space and are therefore involved in many human diseases, such as fibrosis, rheumatoid arthritis, tumor angiogenesis, and metastasis, as well as viral infections. However, the exact mechanism that leads to APN-driven MMP activation is unclear. It was previously shown that extracellular 14-3-3 adapter proteins bind to APN and thereby induce the transcription of MMPs. As a first step, we sought to identify potential 14-3-3–binding sites in the APN sequence. We constructed a set of phosphorylated peptides derived from APN to probe for interactions. We identified and characterized a canonical 14-3-3–binding site (site 1) within the flexible, structurally unresolved N-terminal APN region using direct binding fluorescence polarization assays and thermodynamic analysis. In addition, we identified a secondary, noncanonical binding site (site 2), which enhances the binding affinity in combination with site 1 by many orders of magnitude. Finally, we solved crystal structures of 14-3-3σ bound to mono- and bis-phosphorylated APN-derived peptides, which revealed atomic details of the binding mode of mono- and bivalent 14-3-3 interactions. Therefore, our findings shed some light on the first steps of APN-mediated MMP activation and open the field for further investigation of this important signaling pathway. American Society for Biochemistry and Molecular Biology 2021-01-13 /pmc/articles/PMC7939381/ /pubmed/33109610 http://dx.doi.org/10.1074/jbc.RA120.014708 Text en © 2020 © 2020 Kiehstaller et al. https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Signal Transduction Kiehstaller, Sebastian Ottmann, Christian Hennig, Sven MMP activation–associated aminopeptidase N reveals a bivalent 14-3-3 binding motif |
title | MMP activation–associated aminopeptidase N reveals a bivalent 14-3-3 binding motif |
title_full | MMP activation–associated aminopeptidase N reveals a bivalent 14-3-3 binding motif |
title_fullStr | MMP activation–associated aminopeptidase N reveals a bivalent 14-3-3 binding motif |
title_full_unstemmed | MMP activation–associated aminopeptidase N reveals a bivalent 14-3-3 binding motif |
title_short | MMP activation–associated aminopeptidase N reveals a bivalent 14-3-3 binding motif |
title_sort | mmp activation–associated aminopeptidase n reveals a bivalent 14-3-3 binding motif |
topic | Signal Transduction |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7939381/ https://www.ncbi.nlm.nih.gov/pubmed/33109610 http://dx.doi.org/10.1074/jbc.RA120.014708 |
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