Cargando…
Identification of invariant chain CD74 as a functional receptor of tissue inhibitor of metalloproteinases-1 (TIMP-1)
Multifunctionality of tissue inhibitor of metalloproteinases-1 (TIMP-1) comprising antiproteolytic as well as cytokinic activity has been attributed to its N-terminal and C-terminal domains, respectively. The molecular basis of the emerging proinflammatory cytokinic activity of TIMP-1 is still not c...
Autores principales: | , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8429975/ https://www.ncbi.nlm.nih.gov/pubmed/34391782 http://dx.doi.org/10.1016/j.jbc.2021.101072 |
_version_ | 1783750648448155648 |
---|---|
author | Schoeps, Benjamin Eckfeld, Celina Flüter, Laura Keppler, Selina Mishra, Ritu Knolle, Percy Bayerl, Felix Böttcher, Jan Hermann, Chris D. Häußler, Daniel Krüger, Achim |
author_facet | Schoeps, Benjamin Eckfeld, Celina Flüter, Laura Keppler, Selina Mishra, Ritu Knolle, Percy Bayerl, Felix Böttcher, Jan Hermann, Chris D. Häußler, Daniel Krüger, Achim |
author_sort | Schoeps, Benjamin |
collection | PubMed |
description | Multifunctionality of tissue inhibitor of metalloproteinases-1 (TIMP-1) comprising antiproteolytic as well as cytokinic activity has been attributed to its N-terminal and C-terminal domains, respectively. The molecular basis of the emerging proinflammatory cytokinic activity of TIMP-1 is still not completely understood. The cytokine receptor invariant chain (CD74) is involved in many inflammation-associated diseases and is highly expressed by immune cells. CD74 triggers zeta chain–associated protein kinase-70 (ZAP-70) signaling–associated activation upon interaction with its only known ligand, the macrophage migration inhibitory factor. Here, we demonstrate TIMP-1–CD74 interaction by coimmunoprecipitation and confocal microscopy in cells engineered to overexpress CD74. In silico docking in HADDOCK predicted regions of the N-terminal domain of TIMP-1 (N-TIMP-1) to interact with CD74. This was experimentally confirmed by confocal microscopy demonstrating that recombinant N-TIMP-1 lacking the entire C-terminal domain was sufficient to bind CD74. Interaction of TIMP-1 with endogenously expressed CD74 was demonstrated in the Namalwa B lymphoma cell line by dot blot binding assays as well as confocal microscopy. Functionally, we demonstrated that TIMP-1–CD74 interaction triggered intracellular ZAP-70 activation. N-TIMP-1 was sufficient to induce ZAP-70 activation and interference with the cytokine-binding site of CD74 using a synthetic peptide–abrogated TIMP-1-mediated ZAP-70 activation. Altogether, we here identified CD74 as a receptor and mediator of cytokinic TIMP-1 activity and revealed TIMP-1 as moonlighting protein harboring both cytokinic and antiproteolytic activity within its N-terminal domain. Recognition of this functional TIMP-1–CD74 interaction may shed new light on clinical attempts to therapeutically target ligand-induced CD74 activity in cancer and other inflammatory diseases. |
format | Online Article Text |
id | pubmed-8429975 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-84299752021-09-13 Identification of invariant chain CD74 as a functional receptor of tissue inhibitor of metalloproteinases-1 (TIMP-1) Schoeps, Benjamin Eckfeld, Celina Flüter, Laura Keppler, Selina Mishra, Ritu Knolle, Percy Bayerl, Felix Böttcher, Jan Hermann, Chris D. Häußler, Daniel Krüger, Achim J Biol Chem Research Article Multifunctionality of tissue inhibitor of metalloproteinases-1 (TIMP-1) comprising antiproteolytic as well as cytokinic activity has been attributed to its N-terminal and C-terminal domains, respectively. The molecular basis of the emerging proinflammatory cytokinic activity of TIMP-1 is still not completely understood. The cytokine receptor invariant chain (CD74) is involved in many inflammation-associated diseases and is highly expressed by immune cells. CD74 triggers zeta chain–associated protein kinase-70 (ZAP-70) signaling–associated activation upon interaction with its only known ligand, the macrophage migration inhibitory factor. Here, we demonstrate TIMP-1–CD74 interaction by coimmunoprecipitation and confocal microscopy in cells engineered to overexpress CD74. In silico docking in HADDOCK predicted regions of the N-terminal domain of TIMP-1 (N-TIMP-1) to interact with CD74. This was experimentally confirmed by confocal microscopy demonstrating that recombinant N-TIMP-1 lacking the entire C-terminal domain was sufficient to bind CD74. Interaction of TIMP-1 with endogenously expressed CD74 was demonstrated in the Namalwa B lymphoma cell line by dot blot binding assays as well as confocal microscopy. Functionally, we demonstrated that TIMP-1–CD74 interaction triggered intracellular ZAP-70 activation. N-TIMP-1 was sufficient to induce ZAP-70 activation and interference with the cytokine-binding site of CD74 using a synthetic peptide–abrogated TIMP-1-mediated ZAP-70 activation. Altogether, we here identified CD74 as a receptor and mediator of cytokinic TIMP-1 activity and revealed TIMP-1 as moonlighting protein harboring both cytokinic and antiproteolytic activity within its N-terminal domain. Recognition of this functional TIMP-1–CD74 interaction may shed new light on clinical attempts to therapeutically target ligand-induced CD74 activity in cancer and other inflammatory diseases. American Society for Biochemistry and Molecular Biology 2021-08-12 /pmc/articles/PMC8429975/ /pubmed/34391782 http://dx.doi.org/10.1016/j.jbc.2021.101072 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Research Article Schoeps, Benjamin Eckfeld, Celina Flüter, Laura Keppler, Selina Mishra, Ritu Knolle, Percy Bayerl, Felix Böttcher, Jan Hermann, Chris D. Häußler, Daniel Krüger, Achim Identification of invariant chain CD74 as a functional receptor of tissue inhibitor of metalloproteinases-1 (TIMP-1) |
title | Identification of invariant chain CD74 as a functional receptor of tissue inhibitor of metalloproteinases-1 (TIMP-1) |
title_full | Identification of invariant chain CD74 as a functional receptor of tissue inhibitor of metalloproteinases-1 (TIMP-1) |
title_fullStr | Identification of invariant chain CD74 as a functional receptor of tissue inhibitor of metalloproteinases-1 (TIMP-1) |
title_full_unstemmed | Identification of invariant chain CD74 as a functional receptor of tissue inhibitor of metalloproteinases-1 (TIMP-1) |
title_short | Identification of invariant chain CD74 as a functional receptor of tissue inhibitor of metalloproteinases-1 (TIMP-1) |
title_sort | identification of invariant chain cd74 as a functional receptor of tissue inhibitor of metalloproteinases-1 (timp-1) |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8429975/ https://www.ncbi.nlm.nih.gov/pubmed/34391782 http://dx.doi.org/10.1016/j.jbc.2021.101072 |
work_keys_str_mv | AT schoepsbenjamin identificationofinvariantchaincd74asafunctionalreceptoroftissueinhibitorofmetalloproteinases1timp1 AT eckfeldcelina identificationofinvariantchaincd74asafunctionalreceptoroftissueinhibitorofmetalloproteinases1timp1 AT fluterlaura identificationofinvariantchaincd74asafunctionalreceptoroftissueinhibitorofmetalloproteinases1timp1 AT kepplerselina identificationofinvariantchaincd74asafunctionalreceptoroftissueinhibitorofmetalloproteinases1timp1 AT mishraritu identificationofinvariantchaincd74asafunctionalreceptoroftissueinhibitorofmetalloproteinases1timp1 AT knollepercy identificationofinvariantchaincd74asafunctionalreceptoroftissueinhibitorofmetalloproteinases1timp1 AT bayerlfelix identificationofinvariantchaincd74asafunctionalreceptoroftissueinhibitorofmetalloproteinases1timp1 AT bottcherjan identificationofinvariantchaincd74asafunctionalreceptoroftissueinhibitorofmetalloproteinases1timp1 AT hermannchrisd identificationofinvariantchaincd74asafunctionalreceptoroftissueinhibitorofmetalloproteinases1timp1 AT haußlerdaniel identificationofinvariantchaincd74asafunctionalreceptoroftissueinhibitorofmetalloproteinases1timp1 AT krugerachim identificationofinvariantchaincd74asafunctionalreceptoroftissueinhibitorofmetalloproteinases1timp1 |