Citrullination of fibronectin alters integrin clustering and focal adhesion stability promoting stromal cell invasion

Stefanelli, Victoria L.; Choudhury, Shilpa; Hu, Ping; Liu, Yining; Schwenzer, Anja; Yeh, Chiuan-Ren; Chambers, Dwight M.; Pesson, Kelly; Li, Wei; Segura, Tatiana; Midwood, Kim S.; Torres, Matthew P.; Barker, Thomas H.

doi:10.1016/j.matbio.2019.04.002

Cited by 43 publications

(20 citation statements)

References 68 publications

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“…The slow dissociation rate for the outer barrier suggests a relatively high affinity bond between α 5 β 1 -integrin and fibronectin. This is consistent with the cell attachment function of integrin-ECM binding and suggests an integrin interaction through both the RGD and the synergy binding sites of fibronectin ( 30 , 63 , 64 ). Activation results in the change in conformation of the β 1 subunit and permits higher affinity binding with ligands ( 65 ).…”

Section: Discussionsupporting

confidence: 84%

Distinct Binding Interactions of α5β1-Integrin and Proteoglycans with Fibronectin

Kennelly

Qwarnström

et al. 2019

Biophysical Journal

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Dynamic single-molecule force spectroscopy was performed to monitor the unbinding of fibronectin with the proteoglycans syndecan-4 (SDC4) and decorin and to compare this with the unbinding characteristics of a 5 b 1-integrin. A single energy barrier was sufficient to describe the unbinding of both SDC4 and decorin from fibronectin, whereas two barriers were observed for the dissociation of a 5 b 1-integrin from fibronectin. The outer (high-affinity) barriers in the interactions of fibronectin with a 5 b 1-integrin and SDC4 are characterized by larger barrier heights and widths and slower dissociation rates than those of the inner (low-affinity) barriers in the interactions of fibronectin with a 5 b 1-integrin and decorin. These results indicate that SDC4 and (ultimately) a 5 b 1-integrin have the ability to withstand deformation in their interactions with fibronectin, whereas the decorinfibronectin interaction is considerably more brittle.

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Section: Discussionsupporting

confidence: 84%

Distinct Binding Interactions of α5β1-Integrin and Proteoglycans with Fibronectin

Kennelly

Qwarnström

et al. 2019

Biophysical Journal

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“…We also noticed that the PADI2 knockout macrophages were more easily detached during trypsinization compared with the controls. Stefanelli et al showed that protein citrullination could alter focal adhesion stability [ 12 ]. Therefore, we speculated that PADI2 knockout might also impair the macrophage adhesion.…”

Section: Resultsmentioning

confidence: 99%

The Essential Role of Peptidylarginine Deiminases 2 for Cytokines Secretion, Apoptosis, and Cell Adhesion in Macrophage

Tung

Huang

et al. 2020

IJMS

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Objective: The study aims to investigate the functional roles of peptidylarginine deiminase 2 (PADI2) in macrophages. Methods: The clustered regularly interspaced short palindromic repeats (CRISPR)–CRISPR-associated protein-9 nuclease (Cas9) system was used to knockout PADI2 in U937 cells. U937 cells were introduced to differentiate macrophages and were stimulated with lipopolysaccharides (LPS). The protein expression of PADI2, PADI4, and citrullinated proteins were analyzed by Western blotting. The mRNA and protein levels of interleukin 1 beta (IL-1β), IL-6, and tumor necrosis factor-alpha (TNF-α) were analyzed using RT-PCR and ELISA, respectively. Cell apoptosis was analyzed using flow cytometry. Cell adhesion assay was performed using a commercially available fibrinogen-coated plate. Results: PADI2 knockout could markedly suppress the PADI2 protein expression, but not the PADI4 protein expression. PADI2 knockout decreased the protein levels of citrullinated nuclear factor κB (NF-κB) p65, but not those of citrullinated histone 3, resulting in the decreased mRNA expression levels of IL-1β and TNF-α in the U937 cells and IL-1β and IL-6 in the differentiated macrophages and the macrophages stimulated with LPS. The cytokines levels of IL-1β, IL-6, and TNF-α were all dramatically decreased in the PADI2 knockout group compared with in the controls. PADI2 knockout prevented macrophages apoptosis via the decreased caspase-3, caspase-2, and caspase-9 activation. PADI2 knockout also impaired macrophages adhesion capacity through the decreased protein levels of focal adhesion kinase (FAK), phospho-FAK, paxillin, phospho-paxillin, and p21-activated kinase 1. Conclusion: This study showed that PADI2 could promote IL-1β, IL-6, and TNF-α production in macrophages, promote macrophage apoptosis through caspase-3, caspase-2, and caspase-9 activation and enhance cell adhesion via FAK, paxillin, and PAK1. Therefore, targeting PADI2 could be used as a novel strategy for controlling inflammation caused by macrophages.

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“…Fibronectin is associated with a number of pathologies, including cancer and fibrosis [113]. Fibronectin has been previously found to be deiminated in various sites, which has been related to autoimmunity [114], and also found to support wound healing [115]. In birds, fibronectin is an acute phase protein in chickens (Gallus gallus), responding to infection and changes in temperature [67].…”

Section: Discussionmentioning

confidence: 99%

Protein Deimination and Extracellular Vesicle Profiles in Antarctic Seabirds

Phillips

Kraev

Lange

2020

Biology

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Pelagic seabirds are amongst the most threatened of all avian groups. They face a range of immunological challenges which seem destined to increase due to environmental changes in their breeding and foraging habitats, affecting prey resources and exposure to pollution and pathogens. Therefore, the identification of biomarkers for the assessment of their health status is of considerable importance. Peptidylarginine deiminases (PADs) post-translationally convert arginine into citrulline in target proteins in an irreversible manner. PAD-mediated deimination can cause structural and functional changes in target proteins, allowing for protein moonlighting in physiological and pathophysiological processes. PADs furthermore contribute to the release of extracellular vesicles (EVs), which play important roles in cellular communication. In the present study, post-translationally deiminated protein and EV profiles of plasma were assessed in eight seabird species from the Antarctic, representing two avian orders: Procellariiformes (albatrosses and petrels) and Charadriiformes (waders, auks, gulls and skuas). We report some differences between the species assessed, with the narrowest EV profiles of 50–200 nm in the northern giant petrel Macronectes halli, and the highest abundance of larger 250–500 nm EVs in the brown skua Stercorarius antarcticus. The seabird EVs were positive for phylogenetically conserved EV markers and showed characteristic EV morphology. Post-translational deimination was identified in a range of key plasma proteins critical for immune response and metabolic pathways in three of the bird species under study; the wandering albatross Diomedea exulans, south polar skua Stercorarius maccormicki and northern giant petrel. Some differences in Gene Ontology (GO) biological and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways for deiminated proteins were observed between these three species. This indicates that target proteins for deimination may differ, potentially contributing to a range of physiological functions relating to metabolism and immune response, as well as to key defence mechanisms. PAD protein homologues were identified in the seabird plasma by Western blotting via cross-reaction with human PAD antibodies, at an expected 75 kDa size. This is the first study to profile EVs and to identify deiminated proteins as putative novel plasma biomarkers in Antarctic seabirds. These biomarkers may be further refined to become useful indicators of physiological and immunological status in seabirds—many of which are globally threatened.

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Citrullination of fibronectin alters integrin clustering and focal adhesion stability promoting stromal cell invasion

Cited by 43 publications

References 68 publications

Distinct Binding Interactions of α5β1-Integrin and Proteoglycans with Fibronectin

Distinct Binding Interactions of α5β1-Integrin and Proteoglycans with Fibronectin

The Essential Role of Peptidylarginine Deiminases 2 for Cytokines Secretion, Apoptosis, and Cell Adhesion in Macrophage

Protein Deimination and Extracellular Vesicle Profiles in Antarctic Seabirds

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