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Multivalent Interactions of Human Primary Amine Oxidase with the V and C2(2) Domains of Sialic Acid-Binding Immunoglobulin-Like Lectin-9 Regulate Its Binding and Amine Oxidase Activity
Sialic acid-binding immunoglobulin-like lectin-9 (Siglec-9) on leukocyte surface is a counter-receptor for endothelial cell surface adhesin, human primary amine oxidase (hAOC3), a target protein for anti-inflammatory agents. This interaction can be used to detect inflammation and cancer in vivo, sin...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5125647/ https://www.ncbi.nlm.nih.gov/pubmed/27893774 http://dx.doi.org/10.1371/journal.pone.0166935 |
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author | Elovaara, Heli Parkash, Vimal Fair-Mäkelä, Ruth Salo-Ahen, Outi M. H. Guédez, Gabriela Bligt-Lindén, Eva Grönholm, Janne Jalkanen, Sirpa Salminen, Tiina A. |
author_facet | Elovaara, Heli Parkash, Vimal Fair-Mäkelä, Ruth Salo-Ahen, Outi M. H. Guédez, Gabriela Bligt-Lindén, Eva Grönholm, Janne Jalkanen, Sirpa Salminen, Tiina A. |
author_sort | Elovaara, Heli |
collection | PubMed |
description | Sialic acid-binding immunoglobulin-like lectin-9 (Siglec-9) on leukocyte surface is a counter-receptor for endothelial cell surface adhesin, human primary amine oxidase (hAOC3), a target protein for anti-inflammatory agents. This interaction can be used to detect inflammation and cancer in vivo, since the labeled peptides derived from the second C2 domain (C2(2)) of Siglec-9 specifically bind to the inflammation-inducible hAOC3. As limited knowledge on the interaction between Siglec-9 and hAOC3 has hampered both hAOC3-targeted drug design and in vivo imaging applications, we have now produced and purified the extracellular region of Siglec-9 (Siglec-9-EC) consisting of the V, C2(1) and C2(2) domains, modeled its 3D structure and characterized the hAOC3–Siglec-9 interactions using biophysical methods and activity/inhibition assays. Our results assign individual, previously unknown roles for the V and C2(2) domains. The V domain is responsible for the unusually tight Siglec-9–hAOC3 interactions whereas the intact C2(2) domain of Siglec-9 is required for modulating the enzymatic activity of hAOC3, crucial for the hAOC3-mediated leukocyte trafficking. By characterizing the Siglec-9-EC mutants, we could conclude that R120 in the V domain likely interacts with the terminal sialic acids of hAOC3 attached glycans whereas residues R284 and R290 in C2(2) are involved in the interactions with the active site channel of hAOC3. Furthermore, the C2(2) domain binding enhances the enzymatic activity of hAOC3 although the sialic acid-binding capacity of the V domain of Siglec-9 is abolished by the R120S mutation. To conclude, our results prove that the V and C2(2) domains of Siglec-9-EC interact with hAOC3 in a multifaceted and unique way, forming both glycan-mediated and direct protein-protein interactions, respectively. The reported results on the mechanism of the Siglec-9–hAOC3 interaction are valuable for the development of hAOC3-targeted therapeutics and diagnostic tools. |
format | Online Article Text |
id | pubmed-5125647 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-51256472016-12-15 Multivalent Interactions of Human Primary Amine Oxidase with the V and C2(2) Domains of Sialic Acid-Binding Immunoglobulin-Like Lectin-9 Regulate Its Binding and Amine Oxidase Activity Elovaara, Heli Parkash, Vimal Fair-Mäkelä, Ruth Salo-Ahen, Outi M. H. Guédez, Gabriela Bligt-Lindén, Eva Grönholm, Janne Jalkanen, Sirpa Salminen, Tiina A. PLoS One Research Article Sialic acid-binding immunoglobulin-like lectin-9 (Siglec-9) on leukocyte surface is a counter-receptor for endothelial cell surface adhesin, human primary amine oxidase (hAOC3), a target protein for anti-inflammatory agents. This interaction can be used to detect inflammation and cancer in vivo, since the labeled peptides derived from the second C2 domain (C2(2)) of Siglec-9 specifically bind to the inflammation-inducible hAOC3. As limited knowledge on the interaction between Siglec-9 and hAOC3 has hampered both hAOC3-targeted drug design and in vivo imaging applications, we have now produced and purified the extracellular region of Siglec-9 (Siglec-9-EC) consisting of the V, C2(1) and C2(2) domains, modeled its 3D structure and characterized the hAOC3–Siglec-9 interactions using biophysical methods and activity/inhibition assays. Our results assign individual, previously unknown roles for the V and C2(2) domains. The V domain is responsible for the unusually tight Siglec-9–hAOC3 interactions whereas the intact C2(2) domain of Siglec-9 is required for modulating the enzymatic activity of hAOC3, crucial for the hAOC3-mediated leukocyte trafficking. By characterizing the Siglec-9-EC mutants, we could conclude that R120 in the V domain likely interacts with the terminal sialic acids of hAOC3 attached glycans whereas residues R284 and R290 in C2(2) are involved in the interactions with the active site channel of hAOC3. Furthermore, the C2(2) domain binding enhances the enzymatic activity of hAOC3 although the sialic acid-binding capacity of the V domain of Siglec-9 is abolished by the R120S mutation. To conclude, our results prove that the V and C2(2) domains of Siglec-9-EC interact with hAOC3 in a multifaceted and unique way, forming both glycan-mediated and direct protein-protein interactions, respectively. The reported results on the mechanism of the Siglec-9–hAOC3 interaction are valuable for the development of hAOC3-targeted therapeutics and diagnostic tools. Public Library of Science 2016-11-28 /pmc/articles/PMC5125647/ /pubmed/27893774 http://dx.doi.org/10.1371/journal.pone.0166935 Text en © 2016 Elovaara 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 Elovaara, Heli Parkash, Vimal Fair-Mäkelä, Ruth Salo-Ahen, Outi M. H. Guédez, Gabriela Bligt-Lindén, Eva Grönholm, Janne Jalkanen, Sirpa Salminen, Tiina A. Multivalent Interactions of Human Primary Amine Oxidase with the V and C2(2) Domains of Sialic Acid-Binding Immunoglobulin-Like Lectin-9 Regulate Its Binding and Amine Oxidase Activity |
title | Multivalent Interactions of Human Primary Amine Oxidase with the V and C2(2) Domains of Sialic Acid-Binding Immunoglobulin-Like Lectin-9 Regulate Its Binding and Amine Oxidase Activity |
title_full | Multivalent Interactions of Human Primary Amine Oxidase with the V and C2(2) Domains of Sialic Acid-Binding Immunoglobulin-Like Lectin-9 Regulate Its Binding and Amine Oxidase Activity |
title_fullStr | Multivalent Interactions of Human Primary Amine Oxidase with the V and C2(2) Domains of Sialic Acid-Binding Immunoglobulin-Like Lectin-9 Regulate Its Binding and Amine Oxidase Activity |
title_full_unstemmed | Multivalent Interactions of Human Primary Amine Oxidase with the V and C2(2) Domains of Sialic Acid-Binding Immunoglobulin-Like Lectin-9 Regulate Its Binding and Amine Oxidase Activity |
title_short | Multivalent Interactions of Human Primary Amine Oxidase with the V and C2(2) Domains of Sialic Acid-Binding Immunoglobulin-Like Lectin-9 Regulate Its Binding and Amine Oxidase Activity |
title_sort | multivalent interactions of human primary amine oxidase with the v and c2(2) domains of sialic acid-binding immunoglobulin-like lectin-9 regulate its binding and amine oxidase activity |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5125647/ https://www.ncbi.nlm.nih.gov/pubmed/27893774 http://dx.doi.org/10.1371/journal.pone.0166935 |
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