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Benwell, Christopher J., Johnson, Robert T., Taylor, James A. G. E., Lambert, Jordi and Robinson, Stephen D. 
ORCID: https://orcid.org/0000-0002-6606-7588
(2024)
A proteomics approach to isolating neuropilin-dependent α5 integrin trafficking pathways: Neuropilin 1 and 2 co-traffic α5 integrin through endosomal p120RasGAP to promote polarised fibronectin fibrillogenesis in endothelial cells.
Communications Biology, 7.
ISSN 2399-3642
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Abstract
Integrin trafficking to and from membrane adhesions is a crucial mechanism that dictates many aspects of a cell’s behaviour, including motility, polarisation, and invasion. In endothelial cells (ECs), the intracellular traffic of α5 integrin is regulated by both neuropilin 1 (NRP1) and neuropilin 2 (NRP2), yet the redundancies in function between these co-receptors remain unclear. Moreover, the endocytic complexes that participate in NRP-directed traffic remain poorly annotated. Here we identify an important role for the GTPase-activating protein p120RasGAP in ECs, promoting the recycling of α5 integrin from early endosomes. Mechanistically, p120RasGAP enables transit of endocytosed α5 integrin-NRP1-NRP2 complexes to Rab11+ recycling endosomes, promoting cell polarisation and fibronectin (FN) fibrillogenesis. Silencing of both NRP receptors, or p120RasGAP, resulted in the accumulation of α5 integrin in early endosomes, a loss of α5 integrin from surface adhesions, and attenuated EC polarisation. Endothelial-specific deletion of both NRP1 and NRP2 in the postnatal retina recapitulated our in vitro findings, severely impairing FN fibrillogenesis and polarised sprouting. Our data assign an essential role for p120RasGAP during integrin traffic in ECs and support a hypothesis that NRP receptors co-traffic internalised cargoes. Importantly, we utilise comparative proteomics analyses to isolate a comprehensive map of NRP1-dependent and NRP2-dependent α5 integrin interactions in ECs.
| Item Type: | Article |
|---|---|
| Additional Information: | Data availability statement: The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation, to any qualified researcher. The source data behind the graphs in the paper can be found in Supplementary Data 1. Uncropped and unedited blot/gel images can be found in Supplementary Figs. 7–9. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE79 partner repository with the dataset identifier PXD051778. Funding Information: This work was supported by funding from BHF (grant number PG/22/11033); S.D.R. gratefully acknowledges the support of the Biotechnology and Biological Sciences Research Council (BBSRC); this research was partially funded by the BBSRC Institute Strategic Programme Food Microbiome and Health BB/X011054/1 and its constituent project BBS/E/F/000PR13632. |
| Uncontrolled Keywords: | medicine (miscellaneous),biochemistry, genetics and molecular biology(all),agricultural and biological sciences(all) ,/dk/atira/pure/subjectarea/asjc/2700/2701 |
| Faculty \ School: | Faculty of Science > School of Biological Sciences |
| UEA Research Groups: | Faculty of Medicine and Health Sciences > Research Centres > Norwich Institute for Healthy Aging Faculty of Science > Research Groups > Cells and Tissues |
| Related URLs: | |
| Depositing User: | LivePure Connector |
| Date Deposited: | 23 Oct 2024 09:30 |
| Last Modified: | 01 Nov 2024 00:53 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/97121 |
| DOI: | 10.1038/s42003-024-06320-4 |
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