A proteomics-based survey reveals thrombospondin-4 as a ligand regulated by the mannose receptor in the injured lung

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Receptor-mediated cellular uptake of specific ligands constitutes an important step in the dynamic regulation of individual protein levels in extracellular fluids. With a focus on the inflammatory lung, we here performed a proteomics-based search for novel ligands regulated by the mannose receptor (MR), a macrophage-expressed endocytic receptor. WT and MR-deficient mice were exposed to lipopolysaccharide, after which the protein content in their lung epithelial lining fluid was compared by tandem mass tag-based mass spectrometry. More than 1200 proteins were identified in the epithelial lining fluid using this unbiased approach, but only six showed a statistically different abundance. Among these, an unexpected potential new ligand, thrombospondin-4 (TSP-4), displayed a striking 17-fold increased abundance in the MR-deficient mice. Experiments using exogenous addition of TSP-4 to MR-transfected CHO cells or MR-positive alveolar macrophages confirmed that TSP-4 is a ligand for MR-dependent endocytosis. Similar studies revealed that the molecular interaction with TSP-4 depends on both the lectin activity and the fibronectin type-II domain of MR and that a closely related member of the TSP family, TSP-5, is also efficiently internalized by the receptor. This was unlike the other members of this protein family, including TSPs −1 and −2, which are ligands for a close MR homologue known as urokinase plasminogen activator receptor-associated protein. Our study shows that MR takes part in the regulation of TSP-4, an important inflammatory component in the injured lung, and that two closely related endocytic receptors, expressed on different cell types, undertake the selective endocytosis of distinct members of the TSP family.

OriginalsprogEngelsk
Artikelnummer107284
TidsskriftJournal of Biological Chemistry
Vol/bind300
Udgave nummer5
Antal sider13
ISSN0021-9258
DOI
StatusUdgivet - 2024

Bibliografisk note

Funding Information:
We thank Katharina Hassing, Britt Kongstofte, Mia Kristine Gr\u00F8nning H\u00F8g, and Lene H. Blicher for expert technical assistance. Mass spectrometry analysis was performed at the DTU Proteomics Core, Technical University of Denmark. K. S. N. H. J. J. S. S. H. E. M. S. and N. B. investigation; K. S. N. H. J. J. H. G. E. M. S. and N. B. methodology; K. S. N. H. J. J. and E. M. S. formal analysis; K. S. N. H. J. J. and E. M. S. validation; K. S. N. and H. J. J. visualization; K. S. N. and N. B. conceptualization; K. S. N. and N. B. writing\u2013original draft; H. J. J. T. H. B. L. H. E. and N. B. funding acquisition; H. J. J. H. G. T. H. B. E. M. S. L. H. E. and N. B. writing\u2013review and editing; T. H. B. L. H. E. and N. B. supervision; N. B. project administration. This study was supported by the Danish Cancer Society (Grant nos. R146-A9326-16-S2, R231-A13820 and R368-A21555 (to N. B.), R231-A13832 and R352-A20542 (to L. H. E.) and R90-A5989 (to H. J. J.); The Novo Nordisk Foundation (Grant Nos. NNF19OC0058603 and NNF22OC0080995 (to N. B.)); the Danish Medical Research Council (Grant No. 4092-00387B (to H. J. J.); the European Union's Horizon 2020 research and innovation programme under the Marie Sk\u0142odowska Curie programme (grant agreement No 801481 (to L. H. E.)); Simon Fougner Hartmann\u00B4s Family Foundation (to N. B. and L. H. E.); the Research Fund of the Danish Capital Region (to N. B.); Innovation Fund Denmark (grant agreement Nos. 2070-00019B and 3140-00034B (to L. H. E.)); the NEYE Foundation (to N. B.); the Intramural Research Program at the National Institute of Dental and Craniofacial Research, National Institutes of Health (Z01DE0699 (to T. H. B.)); and the National Institute of Dental and Craniofacial Research Veterinary Resources Core (ZIC DE000740-05 (to T. H. B.)). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

Funding Information:
This study was supported by the Danish Cancer Society (Grant nos. R146-A9326-16-S2, R231-A13820 and R368-A21555 (to NB), R231-A13832 and R352-A20542 (to LHE) and R90-A5989 (to HJJ); The Novo Nordisk Foundation (Grant Nos. NNF19OC0058603 and NNF22OC0080995 (to NB)); the Danish Medical Research Council (Grant No. 4092-00387B (to HJJ ); the European Union\u2019s Horizon 2020 research and innovation programme under the Marie Sk\u0142odowska Curie programme (grant agreement No 801481 (to LHE)); Simon Fougner Hartmann\u00B4s Family Foundation (to NB and LHE); the Research Fund of the Danish Capital Region (to NB); Innovation Fund Denmark (grant agreement Nos. 2070-00019B and 3140-00034B (to LHE)); the NEYE Foundation (to NB); the Intramural Research Program at the National Institute of Dental and Craniofacial Research, National Institutes of Health (Z01DE0699 (to THB)); and the National Institute of Dental and Craniofacial Research Veterinary Resources Core (ZIC DE000740-05 (to THB)).

Publisher Copyright:
© 2024 The Authors

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