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Biophysical Journal Oct 2022The adaptability of proteins to their work environments is fundamental for cellular life. Here, we describe how the hemopexin-like domain of the multifunctional blood...
The adaptability of proteins to their work environments is fundamental for cellular life. Here, we describe how the hemopexin-like domain of the multifunctional blood glycoprotein vitronectin binds Ca to adapt to excursions of temperature and shear stress. Using X-ray crystallography, molecular dynamics simulations, NMR, and differential scanning fluorimetry, we describe how Ca and its flexible hydration shell enable the protein to perform conformational changes that relay beyond the calcium-binding site and alter the number of polar contacts to enhance conformational stability. By means of mutagenesis, we identify key residues that cooperate with Ca to promote protein stability, and we show that calcium association confers protection against shear stress, a property that is advantageous for proteins that circulate in the vasculature, like vitronectin.
Topics: Calcium; Vitronectin; Protein Binding; Hemopexin; Binding Sites; Crystallography, X-Ray; Protein Conformation
PubMed: 36056555
DOI: 10.1016/j.bpj.2022.08.044 -
Molecules (Basel, Switzerland) Feb 2021Acute coronary syndrome (ACS) is a condition in which the coronary artery supplying blood to the heart is infarcted via formation of a plaque and thrombus, resulting in...
Acute coronary syndrome (ACS) is a condition in which the coronary artery supplying blood to the heart is infarcted via formation of a plaque and thrombus, resulting in abnormal blood supply and high mortality and morbidity. Therefore, the prompt and efficient diagnosis of ACS and the need for new ACS diagnostic biomarkers are important. In this study, we aimed to identify new ACS diagnostic biomarkers with high sensitivity and specificity using a proteomic approach. A discovery set with samples from 20 patients with ACS and 20 healthy controls was analyzed using mass spectrometry. Among the proteins identified, those showing a significant difference between each group were selected. Functional analysis of these proteins was conducted to confirm their association with functions in the diseased state. To determine ACS diagnostic biomarkers, standard peptides of the selected protein candidates from the discovery set were quantified, and these protein candidates were validated in a validation set consisting of the sera of 50 patients with ACS and 50 healthy controls. We showed that hemopexin, leucine-rich α-2-glycoprotein, and vitronectin levels were upregulated, whereas fibronectin level was downregulated, in patients with ACS. Thus, the use of these biomarkers may increase the accuracy of ACS diagnosis.
Topics: Acute Coronary Syndrome; Aged; Biomarkers; Female; Fibronectins; Glycoproteins; Hemopexin; Humans; Male; Mass Spectrometry; Middle Aged; Proteomics; Vitronectin
PubMed: 33672727
DOI: 10.3390/molecules26041136 -
Journal of Thrombosis and Haemostasis :... Oct 2022Severe COVID-19 disease is associated with thrombotic complications and extensive fibrin deposition. This study investigates whether the hemostatic complications in...
BACKGROUND
Severe COVID-19 disease is associated with thrombotic complications and extensive fibrin deposition. This study investigates whether the hemostatic complications in COVID-19 disease arise due to dysregulation of the fibrinolytic system.
METHODS
This prospective study analyzed fibrinolytic profiles of 113 patients hospitalized with COVID-19 disease with 24 patients with non-COVID-19 respiratory infection and healthy controls. Antigens were quantified by Ella system or ELISA, clot lysis by turbidimetric assay, and plasminogen activator inhibitor-1 (PAI-1)/plasmin activity using chromogenic substrates. Clot structure was visualized by confocal microscopy.
RESULTS
PAI-1 and its cofactor, vitronectin, are significantly elevated in patients with COVID-19 disease compared with those with non-COVID-19 respiratory infection and healthy control groups. Thrombin activatable fibrinolysis inhibitor and tissue plasminogen activator were elevated in patients with COVID-19 disease relative to healthy controls. PAI-1 and tissue plasminogen activator (tPA) were associated with more severe COVID-19 disease severity. Clots formed from COVID-19 plasma demonstrate an altered fibrin network, with attenuated fiber length and increased branching. Functional studies reveal that plasmin generation and clot lysis were markedly attenuated in COVID-19 disease, while PAI-1 activity was elevated. Clot lysis time significantly correlated with PAI-1 levels. Stratification of COVID-19 samples according to PAI-1 levels reveals significantly faster lysis when using the PAI-1 resistant (tPA) variant, tenecteplase, over alteplase lysis.
CONCLUSION
This study shows that the suboptimal fibrinolytic response in COVID-19 disease is directly attributable to elevated levels of PAI-1, which attenuate plasmin generation. These data highlight the important prognostic potential of PAI-1 and the possibility of using pre-existing drugs, such as tenecteplase, to treat COVID-19 disease and potentially other respiratory diseases.
Topics: Carboxypeptidase B2; Chromogenic Compounds; Fibrin; Fibrinolysin; Fibrinolysis; Hemostatics; Humans; Plasminogen Activator Inhibitor 1; Prospective Studies; Tenecteplase; Thrombosis; Tissue Plasminogen Activator; Vitronectin; COVID-19 Drug Treatment
PubMed: 35780481
DOI: 10.1111/jth.15806 -
Journal of Clinical Periodontology Aug 2022This study investigated whether a vitronectin-derived peptide (VnP-16) prevents and/or reverses alveolar bone resorption induced by ligature-induced periodontitis in...
AIM
This study investigated whether a vitronectin-derived peptide (VnP-16) prevents and/or reverses alveolar bone resorption induced by ligature-induced periodontitis in rodents and identified the underlying mechanism.
MATERIALS AND METHODS
We evaluated the effects of VnP-16 on osteogenic differentiation in human periodontal ligament cells (hPDLCs), lipopolysaccharide-induced inflammatory responses in gingival fibroblasts, and immune response in T lymphocytes. Ligature-induced periodontitis was induced by ligating the bilateral mandibular first molars for 14 days in rats and for 7 days in mice (n = 10/group). VnP-16 (100 μg/10 μl) was applied topically into the gingival sulcus of rats via intra-sulcular injection, whereas the peptide (50 μg/5 μl) was administered directly into the gingiva of mice via intra-gingival injection. To evaluate the preventive and therapeutic effects of VnP-16, micro-computed tomography analysis and histological staining were then performed.
RESULTS
VnP-16 promoted osteogenic differentiation of periodontal ligament cells and inhibited the production of lipopolysaccharide-induced inflammatory mediators in gingival fibroblasts. Concomitantly, VnP-16 modulated the host immune response by reducing the number of receptor activator of NF-κB ligand (RANKL)-expressing lipopolysaccharide-stimulated CD4 and CD8 T cells, and by suppressing RANKL and interleukin (IL)-17A production. Furthermore, local administration of VnP-16 in rats and mice significantly prevented and reversed alveolar bone loss induced by ligature-induced periodontitis. VnP-16 enhanced osteoblastogenesis and simultaneously inhibited osteoclastogenesis and suppressed RANKL and IL-17A expression in vivo.
CONCLUSIONS
Our findings suggest that VnP-16 acts as a potent therapeutic agent for preventing and treating periodontitis by regulating bone re-modelling and immune and inflammatory responses.
Topics: Alveolar Bone Loss; Animals; CD8-Positive T-Lymphocytes; Humans; Interleukin-17; Ligands; Lipopolysaccharides; Mice; NF-kappa B; Osteogenesis; Periodontitis; RANK Ligand; Rats; Vitronectin; X-Ray Microtomography
PubMed: 35634689
DOI: 10.1111/jcpe.13671 -
Journal of the Neurological Sciences Dec 2016Vitronectin is an extracellular matrix protein, the synthesis of which by glioma cells correlates with tumor grade. The current study was designed to investigate the... (Observational Study)
Observational Study
OBJECTIVE
Vitronectin is an extracellular matrix protein, the synthesis of which by glioma cells correlates with tumor grade. The current study was designed to investigate the relationship between serum vitronectin levels and clinicopathological characteristics, diagnosis and prognosis in glioma patients.
METHODS
In a prospective observatory study, a total of 98 glioma patients, 98 healthy controls, 98 other non-glioma brain tumors, and 98 other non-tumor neurological diseases were recruited. Following univariate analyses, multivariate analyses were performed to explore the associations of serum vitronectin levels with survival and clinicopathological parameters. Receiver operating characteristic curve analysis was done to assess its diagnostic and prognostic predictive value.
RESULTS
Serum vitronectin levels were significantly elevated in glioma patients as compared with other groups. High Wealth Health Organization grade was independently associated with high vitronectin levels. Serum vitronectin levels could significantly distinguish glioma patients from other groups and discriminate high-grade glioma from low-grade glioma. Vitronectin levels markedly predicted 5-year progression and 5-year mortality. Moreover, serum vitronectin was identified as an independent predictor for 5-year overall survival and 5-year progression-free survival as well as 5-year mortality and 5-year progression.
CONCLUSION
Serum vitronectin may be a promising diagnostic and prognostic biomarker that can be detected in the peripheral blood of patients with glioma.
Topics: Adult; Age Factors; Aged; Biomarkers, Tumor; Brain Neoplasms; Disease Progression; Female; Glioma; Humans; Male; Middle Aged; Neoplasm Grading; Prognosis; Prospective Studies; Survival Analysis; Treatment Outcome; Tumor Burden; Vitronectin
PubMed: 27871448
DOI: 10.1016/j.jns.2016.10.022 -
Acta Biomaterialia Dec 2015This study is focused on understanding the underlying mechanisms involved in the improved in vitro and in vivo responses of osteoblasts on poly(sodium styrene sulfonate)...
UNLABELLED
This study is focused on understanding the underlying mechanisms involved in the improved in vitro and in vivo responses of osteoblasts on poly(sodium styrene sulfonate) (poly(NaSS)) functionalized Ti6Al4V surfaces. We probed the contribution of cell-adhesive glycoproteins fibronectin (Fn) and vitronectin (Vn) in the initial adhesion of MC3T3-E1 osteoblastic cells to poly(NaSS) functionalized and control Ti6Al4V surfaces. Firstly, culture media containing serum depleted of Fn and Vn (DD) were used to establish the contribution of Fn and Vn in the adhesion and spreading of cells on poly(NaSS) grafted and control surfaces. Compared to ungrafted surfaces, poly(NaSS) grafted surfaces enhanced the levels of cell adhesion, cell spreading and the formation of intracellular actin cytoskeleton and focal contacts in serum treatments where Fn or Vn were present (FBS, DD+Fn, DD+Vn). Cell responses to Fn were more significant than to Vn. Secondly, blocking Fn and Vn integrin receptors using antibodies to α5β1 (Fn) and αvβ1 (Vn) showed that adhesion of cells to poly(NaSS) grafted surfaces principally involved the Fn integrin receptor α5β1. Thirdly, blocking of the heparin and cell-binding regions of Fn molecule (RGD, C-HB, N-HB) showed that grafting with poly(NaSS) altered the conformation of Fn. Together these outcomes explained why the presence of sulfonate (SO3(-)) groups grafted on the Ti6Al4V surface enhanced the early cell adhesion and spreading processes which determine clinical success for applications that require osseointegration.
STATEMENT OF SIGNIFICANCE
This study is devoted to the basic analysis of the mechanism at the origin of the improved in vitro and in vivo osteoblast cell responses exhibited by poly(sodium styrene sulfonate) (poly(NaSS)) functionalized Ti6Al4V surfaces. The aim was to probe the contribution of cell adhesive glycoproteins fibronectin and vitronectin in the initial adhesion of MC3T3-E1 osteoblastic cells to poly(NaSS) functionalized Ti6Al4V surfaces. The outcomes of this research explained why the presence of SO3(-) (sulfonate) groups grafted on the Ti6Al4V surface enhanced the early cell adhesion and spreading processes which determine clinical success for applications that require osseointegration. This work is a step further in the research of poly(NaSS), a very promising bioactive polymer with potential to the orthopedic and dental fields.
Topics: 3T3 Cells; Alloys; Animals; Cell Adhesion; Fibronectins; Integrins; Mice; Osteoblasts; Surface Properties; Titanium; Vitronectin
PubMed: 26415777
DOI: 10.1016/j.actbio.2015.09.030 -
Biotechnology Journal Feb 2018The term "cell adhesion" represents cell-cell interactions and the interaction between the cell and the extracellular matrix (ECM). These interactions are crucial for... (Review)
Review
The term "cell adhesion" represents cell-cell interactions and the interaction between the cell and the extracellular matrix (ECM). These interactions are crucial for the development of the stem cells niche to determine stem cell shape. The ECM is considered as a natural niche for cell residence. Cell adhesion molecules (CAMs) enable cell-cell interactions and the interactions between the cell and ECM via various mechanisms, such as trans-interaction and the heterophilic interactions. These interactions promote a broad spectrum of cell signaling that directly or indirectly modulates stem cell proliferation, self-renewal property, adhesion, and multilineage differentiation. Over many years, animal-derived feeder layers and culture components have been used for stem cell culture, which produces stem cells unsuitable for clinical applications in the regenerative medicine. This review briefly describes the stages of the development of stem cell culture toward a defined condition and the drawbacks of using animal-derived culture components. Stem cell niche-derived and ECM-derived adhesion molecules and their detailed impact on stem cell adhesion and functions will be discussed. Efficient and novel adhesion molecules for stem cell culture and differentiation are needed for further large-scale applications in tissue regeneration.
Topics: Animals; Cadherins; Cell Adhesion; Cell Adhesion Molecules; Cell Culture Techniques; Cell Differentiation; Cell Proliferation; Collagen; Extracellular Matrix; Feeder Cells; Fibronectins; Humans; Hyaluronic Acid; Integrins; Laminin; Pluripotent Stem Cells; Stem Cell Niche; Vitronectin
PubMed: 29193872
DOI: 10.1002/biot.201700575 -
Annals of Neurology Mar 2016CADASIL is a genetic paradigm of cerebral small vessel disease caused by NOTCH3 mutations that stereotypically lead to the extracellular deposition of NOTCH3 ectodomain...
OBJECTIVE
CADASIL is a genetic paradigm of cerebral small vessel disease caused by NOTCH3 mutations that stereotypically lead to the extracellular deposition of NOTCH3 ectodomain (Notch3(ECD) ) on the vessels. TIMP3 and vitronectin are 2 extracellular matrix proteins that abnormally accumulate in Notch3(ECD) -containing deposits on brain vessels of mice and patients with CADASIL. Herein, we investigated whether increased levels of TIMP3 and vitronectin are responsible for aspects of CADASIL disease phenotypes.
METHODS
Timp3 and vitronectin expression were genetically reduced in TgNotch3(R169C) mice, a well-established preclinical model of CADASIL. A mouse overexpressing human TIMP3 (TgBAC-TIMP3) was developed. Disease-related phenotypes, including cerebral blood flow (CBF) deficits, white matter lesions, and Notch3(ECD) deposition, were evaluated between 6 and 20 months of age.
RESULTS
CBF responses to neural activity (functional hyperemia), topical application of vasodilators, and decreases in blood pressure (CBF autoregulation) were similarly reduced in TgNotch3(R169C) and TgBAC-TIMP3 mice, and myogenic responses of brain arteries were likewise attenuated. These defects were rescued in TgNotch3(R169C) mice by haploinsufficiency of Timp3, although the number of white matter lesions was unaffected. In contrast, haploinsufficiency or loss of vitronectin in TgNotch3(R169C) mice ameliorated white matter lesions, although CBF responses were unchanged. Amelioration of cerebrovascular reactivity or white matter lesions in these mice was not associated with reduced Notch3(ECD) deposition in brain vessels.
INTERPRETATION
Elevated levels of TIMP3 and vitronectin, acting downstream of Notch3(ECD) deposition, play a role in CADASIL, producing divergent influences on early CBF deficits and later white matter lesions.
Topics: Animals; Brain; CADASIL; Cerebrovascular Circulation; Humans; Infant; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Mice, Transgenic; Tissue Inhibitor of Metalloproteinases; Vitronectin; Tissue Inhibitor of Metalloproteinase-4
PubMed: 26648042
DOI: 10.1002/ana.24573 -
Mediators of Inflammation 2023Vitronectin (VTN) has been reported to trigger cell pyroptosis to aggravate inflammation in our previous study. However, the function of VTN in inflammatory bowel...
OBJECTIVE
Vitronectin (VTN) has been reported to trigger cell pyroptosis to aggravate inflammation in our previous study. However, the function of VTN in inflammatory bowel disease (IBD) remains to be addressed.
METHODS
Real-time PCR and western blotting were performed to analyze VTN-regulated intestinal epithelial cell (IEC) differentiation through ferroptosis, and immunofluorescence (IF), luciferase, and chromatin immunoprecipitation were used to identify whether VTN-modulated ferroptosis is dependent on phosphodiesterase 4 (PDE4)/protein kinase A (PKA)/cyclic adenosine monophosphate-response element-binding protein (CREB) cascade pathway. experiment in mice and a pilot study in patients with IBD were used to confirm inhibition of PDE4-alleviated IECs ferroptosis, leading to cell differentiation during mucosal healing.
RESULTS
Herein, we found that caudal-related homeobox transcription factor 2-mediated IECs differentiation was impaired in response to VTN, which was attributed to enhanced ferroptosis characterized by decreased glutathione peroxidase 4 (GPX4) and solute carrier family 7 member 11 expression. Inhibition of ferroptosis in IECs rescued the inhibitory effect of VTN on cell differentiation. Further analysis showed that VTN triggered phosphorylation of PDE4, leading to inhibit PKA/CREB activation and CREB nuclear translocation, which further reduced GPX4 transactivation. Endogenous PKA interacted with CREB, and this interaction was destroyed in response to VTN stimulation. What is more, overexpression of CREB in CaCO cells overcame the promotion of VTN on ferroptosis. Most importantly, inhibition of PDE4 by roflumilast or dipyridamole could alleviate dextran sulfate sodium-induced colitis in mice and in a pilot clinical study confirmed by IF.
CONCLUSIONS
These findings demonstrated that highly expressed VTN disrupted IECs differentiation through PDE4-mediated ferroptosis in IBD, suggesting targeting PDE4 could be a promising therapeutic strategy for patients with IBD.
Topics: Mice; Animals; Vitronectin; Cyclic Nucleotide Phosphodiesterases, Type 4; Ferroptosis; Pilot Projects; Inflammatory Bowel Diseases; Cell Differentiation
PubMed: 37501933
DOI: 10.1155/2023/6623329 -
Clinical Laboratory Jul 2019This study aimed to evaluate the relationship of serum vitronectin and integrin alpha V beta 3 (αvβ3) levels with various clinicopathological parameters of breast...
BACKGROUND
This study aimed to evaluate the relationship of serum vitronectin and integrin alpha V beta 3 (αvβ3) levels with various clinicopathological parameters of breast cancer and to assess the diagnostic value of these markers alone or in combination with the conventional breast cancer biomarker CA15.3.
METHODS
This study included 50 early diagnosed stage I - II primary breast cancer patients, 20 patients with fibroadenoma benign lesions, and 20 apparently normal healthy controls. Integrin αVβ3, vitronectin, and CA15.3 levels were measured using ELISA technique.
RESULTS
Serum levels of integrin αVβ3 and vitronectin were significantly higher in the malignant group than those in the benign group and the control group with (p < 0.001). Significant positive correlation between integrin αvβ3 and vitronectin concentrations was found. Both markers showed significant statistically difference with lymph node, histological grade, tumor stage, and tumor size (p < 0.05). Integrin αvβ3 exhibited the highest sensitivity (70%) and specificity (68%), then vitronectin with 67% and 68%, respectively, followed by CA15.3 showing the least sensitivity and specificity (65% and 62%, respectively). All assessed parameters revealed comparable area under the receiver-operating characteristic curve (AUC) 95% confidence interval (CI) range of 0.581 - 0.822.
CONCLUSIONS
Integrin αvβ3 is a promising biomarker alone or in combination with vitronectin and CA15.3 for diagnosis and prognosis of early stage breast cancer.
Topics: Adult; Biomarkers, Tumor; Breast Neoplasms; Early Detection of Cancer; Female; Humans; Integrin alphaVbeta3; Middle Aged; Mucin-1; Prognosis; ROC Curve; Vitronectin
PubMed: 31307158
DOI: 10.7754/Clin.Lab.2019.181219