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Cells Dec 2022It has been four decades since protein S-glutathionylation was proposed to serve as a regulator of cell metabolism. Since then, this redox-sensitive covalent... (Review)
Review
BACKGROUND
It has been four decades since protein S-glutathionylation was proposed to serve as a regulator of cell metabolism. Since then, this redox-sensitive covalent modification has been identified as a cell-wide signaling platform required for embryonic development and regulation of many physiological functions.
SCOPE OF THE REVIEW
Mitochondria use hydrogen peroxide (HO) as a second messenger, but its availability must be controlled to prevent oxidative distress and promote changes in cell behavior in response to stimuli. Experimental data favor the function of protein S-glutathionylation as a feedback loop for the inhibition of mitochondrial HO production.
MAJOR CONCLUSIONS
The glutathione pool redox state is linked to the availability of HO, making glutathionylation an ideal mechanism for preventing oxidative distress whilst playing a part in desensitizing mitochondrial redox signals.
GENERAL SIGNIFICANCE
The biological significance of glutathionylation is rooted in redox status communication. The present review critically evaluates the experimental evidence supporting its role in negating mitochondrial HO production for cell signaling and prevention of electrophilic stress.
Topics: Hydrogen Peroxide; Protein S; Mitochondria; Glutathione; Oxidation-Reduction
PubMed: 36611901
DOI: 10.3390/cells12010107 -
Pediatrics and Neonatology Oct 2017Regular blood transfusion and compliance with iron chelation therapy has markedly improved life expectancy in thalassemia; however, this improvement is accompanied by...
BACKGROUND
Regular blood transfusion and compliance with iron chelation therapy has markedly improved life expectancy in thalassemia; however, this improvement is accompanied by several complications of this chronic disease including thromboembolic disorders. The objective of this work is to study natural coagulation inhibition as well as the fibrinolysis processes in thalassemic children who are otherwise in a steady state with no overt clinical manifestations of thromboembolism.
METHODS
In a case-control study design conducted at Sohag University Hospital, Sohag, Egypt, 50 thalassemic children and 20 age- and sex-matched healthy controls were compared as regards prothrombin concentration, international normalized ratio, partial thromboplastin time, protein C, protein S, antithrombin III, D-dimers, and thrombin activatable fibrinolysis inhibitor (TAFI).
RESULTS
When compared to healthy controls, natural coagulation inhibitors (protein C, protein S, and antithrombin-III) were significantly lower in thalassemic children (p < 0.0001). While D-dimers showed a significant increase in thalassemic children, TAFI was significantly lower (p < 0.0001). Splenectomized thalassemic children showed significantly lower levels of protein C, protein S and TAFI (p < 0.001, p < 0.0001, p < 0.0001, respectively) when compared to nonsplenectomized thalassemic children.
CONCLUSION
Significant changes in natural coagulation inhibition and fibrinolysis processes favoring thromboembolism can be detected in otherwise healthy thalassemic children. Because these changes are more pronounced in splenectomized patients, study of primary prophylactic strategies in this subgroup is warranted.
Topics: Adolescent; Blood Coagulation Disorders; Blood Proteins; Case-Control Studies; Child; Child, Preschool; Female; Fibrin Fibrinogen Degradation Products; Humans; Infant; Male; Protein S; beta-Thalassemia
PubMed: 28351558
DOI: 10.1016/j.pedneo.2016.07.009 -
Clinical and Applied... Nov 2014The increased risk of cardiovascular and cerebrovascular events in patients with migraine remains unexplained. Prothrombotic states are thought to contribute to this...
The increased risk of cardiovascular and cerebrovascular events in patients with migraine remains unexplained. Prothrombotic states are thought to contribute to this increased risk. The present study aimed to compare the prevalence of prothrombotic states in patients with migraine and headache-free controls. We conducted a case-control study to screen for prothrombotic states protein C, protein S (PS), antithrombin III, factor V Leiden, lupus anticoagulant, anticardiolipin, and anti-β2-glycoprotein 1 antibodies in 101 consecutive patients with migraine and 148 controls. An underlying prothrombotic state was encountered in 11.8% of the patients with migraine, PS deficiency being the most common (4.0%). There was no significant difference in the prevalence of prothrombotic states in patients with migraine compared to controls. Traditional prothrombotic states do not seem to have a higher prevalence in patients with migraine compared to controls.
Topics: Adolescent; Adult; Antibodies, Anticardiolipin; Antithrombin III; Blood Coagulation Disorders; Case-Control Studies; Factor V; Female; Humans; Male; Middle Aged; Migraine Disorders; Protein C; Protein S; beta 2-Glycoprotein I
PubMed: 23637003
DOI: 10.1177/1076029613486538 -
European Journal of Obstetrics,... Apr 2024One serious side effect of combined oral contraceptives (COCs) is venous thromboembolism. Reduced activity in activated protein C-related coagulation pathways is...
OBJECTIVE
One serious side effect of combined oral contraceptives (COCs) is venous thromboembolism. Reduced activity in activated protein C-related coagulation pathways is attributable to low protein S activity in one-third of Japanese patients with deep vein thrombosis. Herer, we quantified the behavior of protein S-specific activity in response to dienogest (DNG) and COCs using the protein S-specific activity assay system to explore its potential utility as a thrombosis marker.
STUDY DESIGN
This was a prospective cohort study. Female patients aged 20 - 49 years who were starting drug treatment for endometriosis using DNG or COCs were enrolled. Blood samples were taken before treatment and at the first, third, and sixth months of treatment. To analyze the primary endpoints, changes in total protein S antigen levels, total protein S activity, and protein S-specific activity from baseline to each time point were estimated using a linear mixed-effects model. All statistical analyses were performed in the SAS software version 9.4 (SAS Institute, Cary, NC). A two-sided P < 0.05 was considered statistically significant.
RESULTS
64 patients took DNG and 34 patients took COCs. Protein S-specific activity did not change significantly from baseline in the six months after treatment started in either group. In the DNG group, total protein S activity and total protein S antigen levels increased slightly from baseline levels after the treatment. The means for total protein S activity and total protein S antigen levels in the COC group remained within reference limits, but they both decreased markedly in the first month and stayed low. Protein S-specific activity in four women remaind below the reference limit throughout the whole study period, suggesting they may have potential protein S deficiencies.
CONCLUSION
The effects of DNG on protein S were negligible, though both total protein S activity and antigen levels decreased soon after COC treatment began and remained low. As there was no VTE event during the study, further studies with larger numbers of patients will be needed to confirm that protein S-specific activity can be a surrogate maker of VTE risk.
Topics: Humans; Female; Contraceptives, Oral, Combined; Endometriosis; Prospective Studies; Nandrolone
PubMed: 38340593
DOI: 10.1016/j.ejogrb.2024.01.028 -
MSystems Dec 2022A protein's function depends on functional residues that determine its binding specificity or its catalytic activity, but these residues are typically not considered...
A protein's function depends on functional residues that determine its binding specificity or its catalytic activity, but these residues are typically not considered when annotating a protein's function. To help biologists investigate the functional residues of proteins, we developed two interactive web-based tools, SitesBLAST and Sites on a Tree. Given a protein sequence, SitesBLAST finds homologs that have known functional residues and shows whether the functional residues are conserved. Sites on a Tree shows how functional residues vary across a protein family by showing them on a phylogenetic tree. These tools are available at http://papers.genomics.lbl.gov/sites. For most microbes of interest, a genome sequence is available, but the function of its proteins is not known. Instead, proteins' functions are predicted from their similarity to other protein sequences. Within a protein's sequence, a few key residues are most important for function, such as catalyzing a chemical reaction or determining what it binds. But most function prediction tools do not take these key residues into account. We developed interactive tools for identifying functional residues in a protein sequence by comparing it to proteins with known functional residues. Our tools also make it easy to compare key residues across many similar proteins. This should help biologists check if a protein's function is predicted correctly, or to predict if groups of similar proteins have conserved functions.
Topics: Phylogeny; Computational Biology; Proteins; Amino Acid Sequence; Data Interpretation, Statistical
PubMed: 36374048
DOI: 10.1128/msystems.00705-22 -
Biochemistry and Biophysics Reports Mar 2016The importance of HS in biology and medicine has been widely recognized in recent years, and protein sulfhydration is proposed to mediate the direct actions of HS...
The importance of HS in biology and medicine has been widely recognized in recent years, and protein sulfhydration is proposed to mediate the direct actions of HS bioactivity in the body. Thioredoxin 1 (Trx1) is an important reducing enzyme that cleaves disulfides in proteins and acts as an -denitrosylase. The regulation of Trx1 on protein -sulfhydration is unclear. Here we showed that Trx1 facilitates protein -desulfhydration. Overexpression of Trx1 attenuated the basal level and HS-induced protein -sulfhydration by direct interaction with -sulfhydrated proteins, i.e., glyceraldehyde 3-phosphate dehydrogenase and pyruvate carboxylase. In contrast, knockdown of Trx1 mRNA expression by short interfering RNA or blockage of Trx1 redox activity with PX12 or 2,4-dinitrochlorobenzene enhanced protein -sulfhydration. Mutation of cysteine-32 but not cysteine-35 in the Trp-Cys-Gly-Pro-Cys motif eliminated the binding of Trx1 with -sulfhydrated proteins and abolished the -desulfhydrating effect of Trx1. All these data suggest that Trx1 acts as an -desulfhydrase.
PubMed: 28955804
DOI: 10.1016/j.bbrep.2015.11.012 -
Blood Advances Jan 2022Protein S is a cofactor in the tissue factor pathway inhibitor (TFPI) anticoagulant pathway. It enhances TFPIα-mediated inhibition of factor (F)Xa activity and...
Protein S is a cofactor in the tissue factor pathway inhibitor (TFPI) anticoagulant pathway. It enhances TFPIα-mediated inhibition of factor (F)Xa activity and generation. The enhancement is dependent on a TFPIα-protein S interaction involving TFPIα Kunitz 3 and protein S laminin G-type (LG)-1. C4b binding protein (C4BP), which binds to protein S LG1, almost completely abolishes its TFPI cofactor function. However, neither the amino acids involved in TFPIα enhancement nor the mechanisms underlying the reduced TFPI cofactor function of C4BP-bound protein S are known. To screen for functionally important regions within protein S LG1, we generated 7 variants with inserted N-linked glycosylation attachment sites. Protein S D253T and Q427N/K429T displayed severely reduced TFPI cofactor function while showing normal activated protein C (APC) cofactor function and C4BP binding. Based on these results, we designed 4 protein S variants in which 4 to 6 surface-exposed charged residues were substituted for alanine. One variant, protein S K255A/E257A/D287A/R410A/K423A/E424A, exhibited either abolished or severely reduced TFPI cofactor function in plasma and FXa inhibition assays, both in the presence or absence of FV-short, but retained normal APC cofactor function and high-affinity C4BP binding. The C4BP β-chain was expressed to determine the mechanisms behind the reduced TFPI cofactor function of C4BP-bound protein S. Like C4BP-bound protein S, C4BP β-chain-bound protein S had severely reduced TFPI cofactor function. These results show that protein S Lys255, Glu257, Asp287, Arg410, Lys423, and Glu424 are critical for protein S-mediated enhancement of TFPIα and that binding of the C4BP β-chain blocks this function.
Topics: Complement C4b-Binding Protein; Factor V; Laminin; Lipoproteins; Protein S; Thrombin
PubMed: 34731882
DOI: 10.1182/bloodadvances.2021005382 -
Journal of Acquired Immune Deficiency... Aug 2022HIV-1 infection is associated with multiple procoagulant changes and increased thrombotic risk. Possible mechanisms for this risk include heigthened expression of...
BACKGROUND
HIV-1 infection is associated with multiple procoagulant changes and increased thrombotic risk. Possible mechanisms for this risk include heigthened expression of procoagulant tissue factor (TF) on circulating monocytes, extracellular vesicles, and viral particles and/or acquired deficiency of protein S (PS), a critical cofactor for the anticoagulant protein C (PC). PS deficiency occurs in up to 76% of people living with HIV-1 (PLWH). As increased ex vivo plasma thrombin generation is a strong predictor of mortality, we investigated whether PS and plasma TF are associated with plasma thrombin generation.
METHODS
We analyzed plasma samples from 9 healthy controls, 17 PLWH on first diagnosis (naive), and 13 PLWH on antiretroviral therapy (ART). Plasma thrombin generation, total and free PS, PC, C4b-binding protein, and TF activity were measured.
RESULTS
We determined that the plasma thrombin generation assay is insensitive to PS, because of a lack of PC activation, and developed a modified PS-sensitive assay. Total plasma PS was reduced in 58% of the naive and 38% of the ART-treated PLWH samples and correlated with increased thrombin generation in the modified assay. Conversely, plasma TF was not increased in our patient population, suggesting that it does not significantly contribute to ex vivo plasma thrombin generation.
CONCLUSION
These data suggest that reduced total plasma PS contributes to the thrombotic risk associated with HIV-1 infection and can serve as a prothrombotic biomarker. In addition, our refined thrombin generation assay offers a more sensitive tool to assess the functional consequences of acquired PS deficiency in PLWH.
Topics: Biomarkers; HIV Infections; Humans; Protein S; Thrombin; Thromboplastin
PubMed: 35616596
DOI: 10.1097/QAI.0000000000002994 -
Frontiers in Cardiovascular Medicine 2021Binge drinking has become the most common and deadly pattern of excessive alcohol use in the United States, especially among younger adults. It is closely related to...
Binge drinking has become the most common and deadly pattern of excessive alcohol use in the United States, especially among younger adults. It is closely related to the increased risk of cardiovascular disease. Oxidative stress as a result of ethanol metabolism is the primary pathogenic factor for alcohol-induced end organ injury, but the role of protein S-glutathionylation-a reversible oxidative modification of protein cysteine thiol groups that mediates cellular actions by oxidants-in binge drinking-associated cardiovascular disease has not been explored. The present study defines the effect of alcohol binge drinking on the formation of protein S-glutathionylation in a mouse model of atherosclerosis. To mimic the weekend binge drinking pattern in humans, ApoE deficient ( ) mice on the Lieber-DeCarli liquid diet received ethanol or isocaloric maltose (as a control) gavages (5 g/kg/day, 2 consecutive days/week) for 6 weeks. The primary alcohol-targeted organs (liver, brain), and cardiovascular system (heart, aorta, lung) of these two groups of the mice were determined by measuring the protein S-glutathionylation levels and its regulatory enzymes including [Glutaredoxin1(Grx1), glutathione reductase (GR), glutathione-S-transferase Pi (GST-π)], as well as by assessing aortic endothelial function and liver lipid levels. Our results showed that binge drinking selectively stimulated protein S-glutathionylation in aorta, liver, and brain, which coincided with altered glutathionylation regulatory enzyme expression that is downregulated Grx1 and upregulated GST-π in aorta, massive upregulation of GST-π in liver, and no changes in Grx1 and GST-π in brain. Functionally, binge drinking induced aortic endothelial cell function, as reflected by increased aortic permeability and reduced flow-mediated vasodilation. This study is the first to provide evidence for differential effects of binge drinking on formation of protein S-glutathionylation and its enzymatic regulation system in major alcohol-target organs and cardiovascular system. The selective induction of protein S-glutathionylation in aorta and liver is associated with aortic endothelial dysfunction and fatty liver, which may be a potential redox mechanism for the increased risk of vascular disease in human binge-drinkers.
PubMed: 33796575
DOI: 10.3389/fcvm.2021.649813 -
Blood Coagulation & Fibrinolysis : An... Dec 2019: Protein S Tokushima (p.Lys196Glu) and two protein C gene variants (p.Arg189Trp, p.Lys193del) are hereditary thrombophilia in Japanese and Chinese populations,...
: Protein S Tokushima (p.Lys196Glu) and two protein C gene variants (p.Arg189Trp, p.Lys193del) are hereditary thrombophilia in Japanese and Chinese populations, respectively; however, their diagnosis by plasma analyses is difficult because of the type II deficiency phenotype. Three gene variant genotypes were examined in young Japanese women (n = 231). Plasma total protein S activity and total protein S antigen levels were measured using a total protein S assay system, protein C and protein S activities by clot-based methods, and protein C and free protein S antigen levels by latex agglutination methods. protein S Tokushima (p.Lys196Glu) and protein C p.Lys193del variants were prevalent among participants with allele frequencies of 1.08 and 0.86%, respectively, whereas any carrier of protein C p.Arg189Trp variant was not identified. The plasma phenotype of the type II deficiency of protein S Tokushima heterozygotes was demonstrated by decreased total protein S activity with a normal total protein S antigen level; however, the protein C activities of protein C p.Lys193del heterozygotes were within reference intervals, whereas their protein C antigen levels were elevated. We compared the diagnostic accuracy of the total protein S activity/total protein S antigen ratio for identifying protein S Tokushima heterozygotes with that of the clot-based protein S activity/free protein S antigen ratio and found that sensitivity and specificity of 100% each was only achieved by the former. Protein S Tokushima and protein C p.Lys193del are prevalent among young Japanese women, and a plasma analysis using the total protein S assay system is more accurate than the clot-based protein S activity/free protein S antigen ratio for diagnosing protein S Tokushima carriers.
Topics: Adult; Antigens; Asian People; Female; Gene Frequency; Heterozygote; Humans; Japan; Mutation; Phenotype; Plasma; Protein C; Protein S; Thrombophilia; Young Adult
PubMed: 31490209
DOI: 10.1097/MBC.0000000000000854