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Zhonghua Xue Ye Xue Za Zhi = Zhonghua... Jan 2022To analyze the clinical manifestations and molecular pathogenesis of 18 patients with inherited protein S (PS) deficiency. Eighteen patients with inherited PS...
To analyze the clinical manifestations and molecular pathogenesis of 18 patients with inherited protein S (PS) deficiency. Eighteen patients with inherited PS deficiency who were admitted to the Institute of Hematology & Blood Diseases Hospital from June 2016 to February 2019 were analyzed: activity of protein C (PC) and antithrombin (AT) , PS activity were measured for phenotype diagnosis; high throughput sequencing (HTS) was used for screening of coagulation disease-related genes; Sanger sequencing was used to confirm candidate variants; Swiss-model was used for three-dimensional structure analysis. The PS:C of 18 patients ranged from 12.5 to 48.2 U/dL. Among them, 16 cases developed deep vein thrombosis, including 2 cases each with mesenteric vein thrombosis and cerebral infarction, and 1 case each with pulmonary embolism and deep vein thrombosis during pregnancy. A total of 16 PROS1 gene mutations were detected, and 5 nonsense mutations (c.134_162del/p.Leu45*, c.847G>T/p.Glu283*, c.995_996delAT/p.Tyr332*, c.1359G> A/p.Trp453*, c.1474C>T/p.Gln492*) , 2 frameshift mutations (c.1460delG/p.Gla487Valfs*9 and c.1747_1750delAATC/p.Asn583Wfs*9) and 1 large fragment deletion (exon9 deletion) were reported for the first time. In addition, the PS:C of the deep vein thrombosis during pregnancy case was 55.2 U/dL carrying PROC gene c.565C>T/p.Arg189Trp mutation. The newly discovered gene mutations enriched the PROS1 gene mutation spectrum which associated with inherited PS deficiency.
Topics: Antithrombin III; Female; Genetic Testing; Humans; Mutation; Pregnancy; Protein C; Protein S; Protein S Deficiency
PubMed: 35231993
DOI: 10.3760/cma.j.issn.0253-2727.2022.01.010 -
The Biochemical Journal Sep 2003The irreversible oxidation of cysteine residues can be prevented by protein S-thiolation, a process by which protein SH groups form mixed disulphides with...
The irreversible oxidation of cysteine residues can be prevented by protein S-thiolation, a process by which protein SH groups form mixed disulphides with low-molecular-mass thiols such as glutathione. We report here the target proteins which are modified in yeast cells in response to H(2)O(2). In particular, a range of glycolytic and related enzymes (Tdh3, Eno2, Adh1, Tpi1, Ald6 and Fba1), as well as translation factors (Tef2, Tef5, Nip1 and Rps5) are identified. The oxidative stress conditions used to induce S-thiolation are shown to inhibit GAPDH (glyceraldehyde-3-phosphate dehydrogenase), enolase and alcohol dehydrogenase activities, whereas they have no effect on aldolase, triose phosphate isomerase or aldehyde dehydrogenase activities. The inhibition of GAPDH, enolase and alcohol dehydrogenase is readily reversible once the oxidant is removed. In addition, we show that peroxide stress has little or no effect on glucose-6-phosphate dehydrogenase or 6-phosphogluconate dehydrogenase, the enzymes that catalyse NADPH production via the pentose phosphate pathway. Thus the inhibition of glycolytic flux is proposed to result in glucose equivalents entering the pentose phosphate pathway for the generation of NADPH. Radiolabelling is used to confirm that peroxide stress results in a rapid and reversible inhibition of protein synthesis. Furthermore, we show that glycolytic enzyme activities and protein synthesis are irreversibly inhibited in a mutant that lacks glutathione, and hence cannot modify proteins by S-thiolation. In summary, protein S-thiolation appears to serve an adaptive function during exposure to an oxidative stress by reprogramming metabolism and protecting protein synthesis against irreversible oxidation.
Topics: Alcohol Dehydrogenase; Enzyme Inhibitors; Glucose; Glyceraldehyde-3-Phosphate Dehydrogenases; Glycolysis; Hydrogen Peroxide; Oxidation-Reduction; Oxidative Stress; Pentose Phosphate Pathway; Phosphopyruvate Hydratase; Protein Biosynthesis; Protein S; Saccharomyces cerevisiae Proteins; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Sulfhydryl Compounds
PubMed: 12755685
DOI: 10.1042/BJ20030414 -
Indian Journal of Medical Microbiology 2018Human immunodeficiency virus (HIV) may result in variable haematological manifestations. Thrombotic events are more common among HIV-infected persons than the general...
INTRODUCTION
Human immunodeficiency virus (HIV) may result in variable haematological manifestations. Thrombotic events are more common among HIV-infected persons than the general population, possibly due to the increased inflammatory/hypercoagulable state and presence of concurrent comorbidities.
AIMS AND OBJECTIVES
(1) Screen for coagulation abnormalities in HIV-infected patients. (2) Detect certain prothrombotic factors such as deficiency of protein C and protein S and elevation of homocysteine as possible precursors of coagulation defects in HIV patients. (3) Correlation of coagulation abnormalities with CD4 counts.
METHODS
A pilot study of 1-year duration conducted in the Department of Pathology in collaboration with ART centre, KGMU Lucknow. All diagnosed HIV-seropositive patients (n = 30) who were not taking Vitamin K, antithrombotic and antiplatelet drugs including aspirin, oral contraceptives and not having known protein C/S deficiency were included in the present study as cases. Apart from this, 30 age- and sex-matched healthy individuals were also included in the present study. Assessment of the bleeding time, prothrombin time and activated partial thromboplastin time, complete blood count was done. Protein C and S were measured by calorimetric assay. Serum homocysteine was measured by the semi-automated method. CD4 count was done by flow cytometry.
RESULTS
The findings of the present study suggest a relationship between HIV, its complications and thrombosis. The HIV-seropositive patients have reduced levels of haemoglobin, CD4 counts, platelet counts, mean platelet volume, protein C and S activity as compared to the healthy individuals. Thrombophilic abnormality in the form of hyperhomocysteinaemia is more frequent in HIV-infected patients. All these parameters have a definite correlation with CD4 count.
Topics: Adult; CD4 Lymphocyte Count; Female; HIV Infections; Homocysteine; Humans; Male; Middle Aged; Protein C; Protein S; Thrombosis
PubMed: 30084411
DOI: 10.4103/ijmm.IJMM_15_414 -
Blood Jul 2018There is a Commentary on this article in this issue.
There is a Commentary on this article in this issue.
Topics: Animals; Down-Regulation; Hep G2 Cells; Humans; Hypoxia; Hypoxia-Inducible Factor 1, alpha Subunit; Liver; Mice; Mice, Knockout; Protein S; Thrombosis
PubMed: 29784640
DOI: 10.1182/blood-2018-04-841585 -
Molecular Plant-microbe Interactions :... Apr 2020In orthotospovirus, the nonstructural protein S (NSs) is the RNA-silencing suppressor (RSS) and pathogenicity determinant. Here, we demonstrate that a putative α-helix,...
A Conserved Helix in the C-Terminal Region of Watermelon Silver Mottle Virus Nonstructural Protein S Is Imperative For Protein Stability Affecting Self-Interaction, RNA Silencing Suppression, and Pathogenicity.
In orthotospovirus, the nonstructural protein S (NSs) is the RNA-silencing suppressor (RSS) and pathogenicity determinant. Here, we demonstrate that a putative α-helix, designated H8, spanning amino acids 338 to 369 of the C-terminal region of the NSs protein, is crucial for self-interaction of watermelon silver mottle virus NSs protein and that the H8 affects RSS function. Co-immunoprecipitation, yeast two-hybrid, and bimolecular fluorescence complementation analyses revealed that the triple point mutation (TPM) of H8 amino acids Y338A, H350A, and F353A resulted in NSs protein self-interaction dysfunction. Transient expression of H8-deleted (ΔH8) and TPM NSs proteins in plants by agroinfitration indicated that these proteins have weaker RSS activity and are far less stable than wild-type (WT) NSs. However, an electrophoretic mobility assay revealed that small interfering RNA (siRNA) binding ability of TPM NSs protein is not compromised. The pathogenicity assay of WT NSs protein expressed by the attenuated turnip mosaic virus vector restored severe symptoms in recombinant-infected plants but not for ΔH8 or TPM proteins. Taken together, we conclude that the H8 helix in the C-terminal region of NSs protein is crucial for stabilizing NSs protein through self-interaction to maintain normal functions of RSS and pathogenicity, but not for NSs-siRNA binding activity.
Topics: Protein S; Protein Stability; RNA Interference; Nicotiana; Tospovirus; Viral Nonstructural Proteins; Virulence
PubMed: 31935338
DOI: 10.1094/MPMI-10-19-0279-R -
Redox Biology May 2020The pathogenesis of many human diseases has been attributed to the over production of reactive oxygen species (ROS), particularly superoxide (O) and hydrogen peroxide... (Review)
Review
The pathogenesis of many human diseases has been attributed to the over production of reactive oxygen species (ROS), particularly superoxide (O) and hydrogen peroxide (HO), by-products of metabolism that are generated by the premature reaction of electrons with molecular oxygen (O) before they reach complex IV of the respiratory chain. To date, there are 32 known ROS generators in mammalian cells, 16 of which reside inside mitochondria. Importantly, although these ROS are deleterious at high levels, controlled and temporary bursts in HO production is beneficial to mammalian cells. Mammalian cells use sophisticated systems to take advantage of the second messaging properties of HO. This includes controlling its availability using antioxidant systems and negative feedback loops that inhibit the genesis of ROS at sites of production. At its core, ROS production depends on fuel metabolism. Therefore, desensitizing HO signals would also require the temporary inhibition of fuel combustion and fluxes through metabolic pathways that promote ROS production. Additionally, this would also demand the diversion of fuels and nutrients into pathways that generate NADPH and other molecules required to maintain cellular redox buffering capacity. Therefore, fuel selection and metabolic flux plays an integral role in dictating the strength and duration of cellular redox signals. In the present review I provide an updated view on the function of protein S-glutathionylation, a ubiquitous redox sensitive modification involving the formation of a disulfide between the small molecular antioxidant glutathione and a cysteine residue, in the regulation of cellular metabolism on a global scale. To date, these concepts have mostly been reviewed at the level of mitochondrial bioenergetics in the contexts of health and disease. Careful examination of the literature revealed that glutathionylation is a temporary inhibitor of most metabolic pathways including glycolysis, the Krebs cycle, oxidative phosphorylation, amino acid metabolism, and fatty acid combustion, resulting in the diversion of fuels towards NADPH-producing pathways and the inhibition of ROS production. Armed with this information, I propose that protein S-glutathionylation reactions desensitize HO signals emanating from catabolic pathways using a three-pronged regulatory mechanism; 1) inhibition of metabolic flux through pathways that promote ROS production, 2) diversion of metabolites towards pathways that support antioxidant defenses, and 3) direct inhibition of ROS-generating enzymes.
Topics: Animals; Humans; Hydrogen Peroxide; Oxidation-Reduction; Protein S; Reactive Oxygen Species; Superoxides
PubMed: 32171726
DOI: 10.1016/j.redox.2020.101472 -
Journal of Thrombosis and Haemostasis :... Dec 2019Activated protein C (APC)-mediated inactivation of factor (F)Va is greatly enhanced by protein S. For inactivation to occur, a trimolecular complex among FVa, APC, and...
BACKGROUND
Activated protein C (APC)-mediated inactivation of factor (F)Va is greatly enhanced by protein S. For inactivation to occur, a trimolecular complex among FVa, APC, and protein S must form on the phospholipid membrane. However, direct demonstration of complex formation has proven elusive.
OBJECTIVES
To elucidate the nature of the phospholipid-dependent interactions among APC, protein S, and FVa.
METHODS
We evaluated binding of active site blocked APC to phospholipid-coated magnetic beads in the presence and absence of protein S and/or FVa. The importance of protein S and FV residues were evaluated functionally.
RESULTS
Activated protein C alone bound weakly to phospholipids. Protein S mildly enhanced APC binding to phospholipid surfaces, whereas FVa did not. However, FVa together with protein S enhanced APC binding (>14-fold), demonstrating formation of an APC/protein S/FVa complex. C4b binding protein-bound protein S failed to enhance APC binding, agreeing with its reduced APC cofactor function. Protein S variants (E36A and D95A) with reduced APC cofactor function exhibited essentially normal augmentation of APC binding to phospholipids, but diminished APC/protein S/FVa complex formation, suggesting involvement in interactions dependent upon FVa. Similarly, FVa (W1920R), an APC-resistant FV variant, also did not efficiently incorporate into the trimolecular complex as efficiently as wild-type FVa. FVa inactivation assays suggested that the mutation impairs its affinity for phospholipid membranes and with protein S within the complex.
CONCLUSIONS
FVa plays a central role in the formation of its inactivation complex. Furthermore, membrane proximal interactions among FVa, APC, and protein S are essential for its cofactor function.
Topics: Binding Sites; Blood Coagulation; Calcium-Binding Proteins; Enzyme Activation; Factor Va; HEK293 Cells; Humans; Models, Molecular; Multiprotein Complexes; Phospholipids; Protein Binding; Protein C; Protein Conformation; Protein S; Structure-Activity Relationship; Thrombin; Thromboplastin
PubMed: 31364267
DOI: 10.1111/jth.14594 -
The Journal of Biological Chemistry Jul 1994Inherited resistance to activated protein C (APC) is a recently identified major cause of thrombosis. It is associated with a mutation in the factor V gene affecting one...
Inherited resistance to activated protein C (APC) is a recently identified major cause of thrombosis. It is associated with a mutation in the factor V gene affecting one of the cleavage sites for APC. APC resistance was recently found to be corrected by factor V, suggesting that factor V may have anticoagulant properties as a cofactor to APC. To elucidate this further, we have studied the effect of factor V and protein S, which is a known cofactor to APC, on APC-mediated degradation of factor VIIIa in a purified system. The APC-mediated degradation of factor VIIIa was monitored by a factor X activation reaction using purified factor IXa, phospholipid, and calcium. In the presence of both factor V and protein S, APC was found to inhibit factor VIIIa activity efficiently. APC alone or together with factor V was ineffective, whereas APC in combination with protein S was less efficient than when factor V was also included in the reaction. Two monoclonal antibodies, one against protein S and the other directed toward factor V, were found to inhibit the APC cofactor activity of the factor V-protein S mixture. Factor Va did not express APC cofactor activity, and addition of excess factor Va over factor V did not inhibit the APC cofactor function of a factor V-protein S mixture. In conclusion, the results suggest that factor V and protein S work in synergy as phospholipid-bound cofactors to APC.
Topics: Enzyme Activation; Factor V; Factor VIIIa; Humans; In Vitro Techniques; Macromolecular Substances; Protein C; Protein S
PubMed: 8034625
DOI: No ID Found -
Redox Biology Jun 2023Oxidative stress drives protein S-glutathionylation, which regulates the structure and function of target proteins and is implicated in the pathogenesis of many...
Glutaredoxin 1 protects lens epithelial cells from epithelial-mesenchymal transition by preventing casein kinase 1α S-glutathionylation during posterior capsular opacification.
Oxidative stress drives protein S-glutathionylation, which regulates the structure and function of target proteins and is implicated in the pathogenesis of many diseases. Glutaredoxin 1 (Grx1), a cytoplasmic deglutathionylating enzyme, maintains a reducing environment within the cell under various conditions by reversing S-glutathionylation. Grx1 performs a wide range of antioxidant activities in the lens and prevents protein-thiol mixed disulfide accumulation, reducing protein-protein aggregation, insolubilization, and apoptosis of lens epithelial cells. Oxidative stress is related to epithelial-mesenchymal transition (EMT) during posterior capsular opacification (PCO). However, whether Grx1-regulated protein S-glutathionylation plays an essential role in PCO remains unclear. In this study, we revealed that Grx1 expression was decreased in mice following cataract surgery. Furthermore, the absence of Grx1 elevated oxidative stress and protein S-glutathionylation and aggravated EMT in both in vitro and in vivo models. Concurrently, these results could be reversed by Grx1 overexpression. Notably, liquid chromatography-tandem mass spectrometry results showed that casein kinase 1α (CK1α) was susceptible to S-glutathionylation under oxidative stress, and CK1α S-glutathionylation (CK1α-SSG) was mediated at Cys249. The absence of Grx1 upregulated CK1α-SSG, subsequently decreasing the CK1α-induced phosphorylation of β-catenin at Ser45. The consequential downregulation of degradative β-catenin and upregulation of its nuclear translocation activated the Wnt/β-catenin signaling pathway and aggravated EMT. In conclusion, the downregulated expression of Grx1 in mice following cataract surgery aggravated EMT by upregulating the extent of CK1α-SSG. To the best of our knowledge, our study is the first to report how S-glutathionylation regulates CK1α activity under oxidative stress.
Topics: Animals; Mice; beta Catenin; Casein Kinases; Cataract; Epithelial Cells; Epithelial-Mesenchymal Transition; Glutaredoxins; Glutathione; Protein S
PubMed: 36989576
DOI: 10.1016/j.redox.2023.102676 -
Journal of Thrombosis and Haemostasis :... Feb 2010Protein S and tissue factor pathway inhibitor (TFPI) act together in down-regulating coagulation.
BACKGROUND
Protein S and tissue factor pathway inhibitor (TFPI) act together in down-regulating coagulation.
OBJECTIVE
To investigate the TFPI/protein S system in hereditary and acquired protein S deficiency.
METHODS
Plasma antigen levels of protein S and full-length TFPI were determined in heterozygous type I protein S-deficient individuals (n=35), patients on oral anticoagulant treatment (OAT) (n=29), oral contraceptive (OC) users (n=10) and matched controls. Thrombin generation was determined using calibrated automated thrombography.
RESULTS
Full-length TFPI levels were lower in type I protein S-deficient individuals (76.8+/-33.8%) than in age- and sex-matched controls (128.0+/-59.4%, P<0.001). Among protein S-deficient individuals with thrombosis, those on OAT had not only lower total protein S levels (25.7+/-8.2% vs. 54.7+/-8.2%, P<0.001), but also lower full-length TFPI levels (52.6+/-15.0% vs. 75.4+/-22.9%, P=0.009) than those not on OAT. Similarly, OC users had lower protein S (73.8+/-11.5% vs. 87.9+/-10.8%, P=0.005) and full-length TFPI levels (73.7+/-27.7% vs. 106.4+/-29.2%, P=0.007) than non-users. When triggered with tissue factor, plasma from protein S-deficient individuals generated 3-5-fold more thrombin than control plasma. The difference was only partially corrected by normalization of the protein S level, full correction requiring additional normalization of the TFPI level. Protein S-immunodepletion experiments indicated that free protein S and full-length TFPI form a complex in plasma, and the protein S/TFPI interaction was confirmed by surface plasmon resonance analysis.
CONCLUSIONS
Full-length TFPI binds to protein S in plasma and is reduced in genetic and acquired protein S deficiency. The concomitant TFPI deficiency substantially contributes to the hypercoagulable state associated with protein S deficiency.
Topics: Administration, Oral; Adult; Aged; Anticoagulants; Blood Coagulation; Case-Control Studies; Contraceptives, Oral; Down-Regulation; Female; Genetic Predisposition to Disease; Heterozygote; Humans; Lipoproteins; Male; Middle Aged; Protein Binding; Protein S; Protein S Deficiency; Risk Factors; Thrombin; Thromboplastin; Time Factors; Young Adult
PubMed: 20002538
DOI: 10.1111/j.1538-7836.2009.03712.x