Academic Journal
Regulation of tissue factor activity by interaction with the first PDZ domain of MAGI1
| العنوان: | Regulation of tissue factor activity by interaction with the first PDZ domain of MAGI1 |
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| المؤلفون: | Mohammad, Mohammad A, Featherby, Sophie, Ettelaie, Camille |
| بيانات النشر: | Springer Verlag |
| سنة النشر: | 2024 |
| المجموعة: | University of Hull: Repository@Hull |
| مصطلحات موضوعية: | Tissue factor, MAGI1, PAR2, PDZ domain, De-encryption, Thrombin generation activity, Proximity Ligation Assay, Co-immunoprecipitation |
| الوصف: | Background: Tissue factor (TF) activity is stringently regulated through processes termed encryption. Post-translational modification of TF and its interactions with various protein and lipid moieties allows for a multi-step de-encryption of TF and procoagulant activation. Membrane-associated guanylate kinase-with inverted configuration (MAGI) proteins are known to regulate the localisation and activity of a number of proteins including cell-surface receptors. Methods: The interaction of TF with MAGI1 protein was examined as a means of regulating TF activity. MDA-MB-231 cell line was used which express TF and MAGI1, and respond well to protease activated receptor (PAR)2 activation. Proximity ligation assay (PLA), co-immunoprecipitation and pull-down experiments were used to examine the interaction of TF with MAGI1-3 proteins and to investigate the influence of PAR2 activation. Furthermore, by cloning and expressing the PDZ domains from MAGI1, the TF-binding domain was identified. The ability of the recombinant PDZ domains to act as competitors for MAGI1, allowing the induction of TF procoagulant and signallingactivity was then examined. Results: PLA and fluorescence microscopic analysis indicated that TF predominantly associates with MAGI1 and less with MAGI2 and MAGI3 proteins. The interaction of TF with MAGI1 was also demonstrated by both co-immunoprecipitation of TF with MAGI1, and co-immunoprecipitation of MAGI1 with TF. Moreover, activation of PAR2 resulted in reduction in the association of these two proteins. Pull-down assays using TF-cytoplasmic domain peptides indicated that the phosphorylation of Ser253 within TF prevents its association with MAGI1. Additionally, the five HA-tagged PDZ domains of MAGI1 were overexpressed separately, and the putative TF-binding domain was identified as PDZ1 domain. Expression of this PDZ domain in cells significantly augmented the TF activity measured both as thrombin-generation and also TF-mediated proliferative signalling. Conclusions: Our data indicate a stabilising ... |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| تدمد: | 1477-9560 |
| Relation: | https://hull-repository.worktribe.com/output/4518994; Thrombosis Journal; Volume 22; Issue 1; https://hull-repository.worktribe.com/file/4518994/1/Published%20article |
| DOI: | 10.1186/s12959-023-00580-6 |
| الاتاحة: | https://hull-repository.worktribe.com/file/4518994/1/Published%20article https://hull-repository.worktribe.com/output/4518994 https://doi.org/10.1186/s12959-023-00580-6 |
| Rights: | openAccess ; http://creativecommons.org/licenses/by/4.0 |
| رقم الانضمام: | edsbas.32D434E6 |
| قاعدة البيانات: | BASE |
Full Text Finder | تدمد: | 14779560 |
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| DOI: | 10.1186/s12959-023-00580-6 |