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Journal of the American Chemical Society Aug 2021HS and HO are two redox regulating molecules that play important roles in many physiological and pathological processes. While each of them has distinct biosynthetic...
HS and HO are two redox regulating molecules that play important roles in many physiological and pathological processes. While each of them has distinct biosynthetic pathways and signaling mechanisms, the crosstalk between these two species is also known to cause critical biological responses such as protein S-persulfidation. So far, many chemical tools for the studies of HS and HO have been developed, such as the donors and sensors for HS and HO. However, these tools are normally targeting single species (e.g., only HS or only HO). As such, the crosstalk and synergetic effects between HS and HO have hardly been studied with those tools. In this work, we report a unique HS/HO dual donor system by employing 1-thio-β-d-glucose and glucose oxidase (GOx) as the substrates. This enzymatic system can simultaneously produce HS and HO in a slow and controllable fashion, without generating any bio-unfriendly byproducts. This system was demonstrated to cause efficient S-persulfidation on proteins. In addition, we expanded the system to thiolactose and thioglucose-disulfide; therefore, additional factors (β-galactosidase and cellular reductants) could be introduced to further control the release of HS/HO. This dual release system should be useful for future research on HS and HO.
Topics: Glucose; Glucose Oxidase; Humans; Hydrogen Peroxide; Hydrogen Sulfide; Protein S
PubMed: 34383487
DOI: 10.1021/jacs.1c06372 -
Cureus Oct 2023Protein S is a glycoprotein created by the body that aids in the prevention of a hypercoagulable state. Protein S-deficient patients are placed on anticoagulant...
Protein S is a glycoprotein created by the body that aids in the prevention of a hypercoagulable state. Protein S-deficient patients are placed on anticoagulant regimens, as there is no current definitive cure. Failure to bring balance to the hematological system in these patients will lead to complications such as widespread clot formation and pulmonary embolisms. Here, we present a 74-year-old female who was admitted to the ICU after collapsing. She presented with respiratory failure, urinary tract infection (UTI), and pneumonia. Magnetic resonance imaging (MRI) scans depicted a thrombus in the distal right transverse sinus and sigmoid sinus. Her hematologic workup showed normal levels of homocysteine, fibrinogen, and protein C levels but protein S levels were reduced to 24%. This case displays the intricate presentation of a rare hematological disease as well as the importance of routine follow-up to maintain patient health.
PubMed: 37954832
DOI: 10.7759/cureus.46864 -
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 -
Plant & Cell Physiology Jan 2020Nitric oxide (NO) is a crucial signaling molecule that conveys its bioactivity mainly through protein S-nitrosylation. This is a reversible post-translational...
Nitric oxide (NO) is a crucial signaling molecule that conveys its bioactivity mainly through protein S-nitrosylation. This is a reversible post-translational modification (PTM) that may affect protein function. S-nitrosoglutathione (GSNO) is a cellular NO reservoir and NO donor in protein S-nitrosylation. The enzyme S-nitrosoglutathione reductase (GSNOR) degrades GSNO, thereby regulating indirectly signaling cascades associated with this PTM. Here, the two GSNORs of the legume Lotus japonicus, LjGSNOR1 and LjGSNOR2, have been functionally characterized. The LjGSNOR1 gene is very active in leaves and roots, whereas LjGSNOR2 is highly expressed in nodules. The enzyme activities are regulated in vitro by redox-based PTMs. Reducing conditions and hydrogen sulfide-mediated cysteine persulfidation induced both activities, whereas cysteine oxidation or glutathionylation inhibited them. Ljgsnor1 knockout mutants contained higher levels of S-nitrosothiols. Affinity chromatography and subsequent shotgun proteomics allowed us to identify 19 proteins that are differentially S-nitrosylated in the mutant and the wild-type. These include proteins involved in biotic stress, protein degradation, antioxidant protection and photosynthesis. We propose that, in the mutant plants, deregulated protein S-nitrosylation contributes to developmental alterations, such as growth inhibition, impaired nodulation and delayed flowering and fruiting. Our results highlight the importance of GSNOR function in legume biology.
Topics: Aldehyde Oxidoreductases; Cysteine; Genes, Plant; Lotus; Nitric Oxide; Nitric Oxide Donors; Oxidation-Reduction; Plant Proteins; Protein Processing, Post-Translational; Protein S; Proteomics; S-Nitrosoglutathione; S-Nitrosothiols; Tandem Mass Spectrometry
PubMed: 31529085
DOI: 10.1093/pcp/pcz182 -
Thrombosis Research Mar 2023COVID-19 is associated with an increased thromboembolic risk. However, the mechanisms triggering clot formation in those patients remain unknown.
INTRODUCTION
COVID-19 is associated with an increased thromboembolic risk. However, the mechanisms triggering clot formation in those patients remain unknown.
PATIENTS AND METHODS
In 118 adult Caucasian severe but non-critically ill COVID-19 patients (median age 58 years; 73 % men) and 46 controls, we analyzed in vitro plasma thrombin generation profile (calibrated automated thrombogram [CAT assay]) and investigated thrombophilia-related factors, such as protein C and antithrombin activity, free protein S level, presence of antiphospholipid antibodies and factor V Leiden R506Q and prothrombin G20210A mutations. We also measured circulating von Willebrand factor (vWF) antigen and a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13 (ADAMTS13) antigen and activity. In patients, blood samples were collected on admission to the hospital before starting any therapy, including heparin. Finally, we examined the relationship between observed alterations and disease follow-up, such as thromboembolic complications.
RESULTS
COVID-19 patients showed 17 % lower protein C activity, 22 % decreased free protein S levels, and a higher prevalence of positive results for IgM anticardiolipin antibodies. They also had 151 % increased vWF, and 27 % decreased ADAMTS13 antigens compared with controls (p < 0.001, all). On the contrary, thrombin generation potential was similar to controls. In the follow-up, pulmonary embolism (PE) occurred in thirteen (11 %) patients. They were characterized by a 55 % elevated D-dimer (p = 0.04) and 2.7-fold higher troponin I (p = 0.002) during hospitalization and 29 % shorter time to thrombin peak in CAT assay (p = 0.009) compared to patients without PE.
CONCLUSIONS
In COVID-19, we documented prothrombotic abnormalities of peripheral blood. PE was characterized by more dynamic thrombin generation growth in CAT assay performed on admittance to the hospital.
Topics: Humans; ADAMTS13 Protein; COVID-19; Protein C; Thrombin; von Willebrand Factor; Protein S
PubMed: 36709678
DOI: 10.1016/j.thromres.2023.01.016 -
Inherited Thrombophilia in Chinese CTEPH Patients: A Rather Common Finding in an Uncommon Condition.JACC. Asia Jun 2022
PubMed: 36338408
DOI: 10.1016/j.jacasi.2022.03.005 -
Journal of Agricultural and Food... Feb 2022The objective of the present study was to investigate the regulatory mechanism of protein S-nitrosylation on early postmortem beef muscle apoptosis. Beef semimembranosus...
The objective of the present study was to investigate the regulatory mechanism of protein S-nitrosylation on early postmortem beef muscle apoptosis. Beef semimembranosus (SM) muscles at 45 min postmortem were treated with nitric oxide (NO) donor, control (NaCl solution), or nitric oxide synthase (NOS) inhibitor for 24 h at 4 °C. Bcl-2 expression and mitochondrial membrane potential were significantly increased by the NO donor treatment at 6 h postmortem, while the NOS inhibitor group exhibited a lower Bcl-2 level and mitochondrial membrane potential in comparison with the control ( < 0.05). The cytochrome c expression analysis highlighted that NO donor incubation repressed cytochrome c release from mitochondria to the cytoplasm. Further, S-nitrosylation levels of caspase-3 and caspase-9 were elevated after incubation with the NO donor ( < 0.05), leading to decreased caspase-3 and caspase-9 activities ( < 0.05). The aforementioned findings imply that protein S-nitrosylation mediates postmortem apoptosis of beef SM through the mitochondrial apoptotic pathway.
Topics: Animals; Apoptosis; Caspase 3; Cattle; Cytochromes c; Mitochondria; Nitric Oxide; Protein S
PubMed: 34968404
DOI: 10.1021/acs.jafc.1c06516 -
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 -
Thrombosis Research Feb 2022Protein S is a vitamin K-dependent glycoprotein with important anticoagulant, fibrinolytic, anti-inflammatory, anti-apoptotic, and cytoprotective functions. Congenital...
INTRODUCTION
Protein S is a vitamin K-dependent glycoprotein with important anticoagulant, fibrinolytic, anti-inflammatory, anti-apoptotic, and cytoprotective functions. Congenital protein S deficiency is an autosomal dominant thrombophilia due to protein S gene (PROS1) variations. Our group identified a variation in PROS1 that translates into protein S deficiency: c.50 T > C (p.Leu17Pro). Here, we investigated the mechanisms by which this variation results in protein S deficiency.
MATERIALS AND METHODS
The effect of L17P substitution on protein S signal peptide was predicted by in silico (a computational prediction technique) analysis of hydrophobicity and signal peptide cleavage. Recombinant protein S was overexpressed in HEK293 and COS-7 cells. Intracellular kinetics and extracellular secretion of recombinant protein S-L17P were analyzed by western blotting and immunocytochemistry.
RESULTS
In silico hydrophobicity analysis showed that protein S-L17P had disrupted hydrophobic status in the h-region of its signal peptide. Under normal culture conditions, recombinant protein S -L17P was not detected in either transfectant cell lysates or medium. Upon treatment with a proteasome inhibitor, recombinant protein S-L17P was clearly detected in the cell lysate, but not in the culture medium. Recombinant protein S-L17P did not undergo post-translational modification with N-glycosylation, suggesting that the nascent polypeptide of recombinant protein S-L17P is not transported to the endoplasmic reticulum lumen, but is mislocalized to the cytosol.
CONCLUSION
PROS1-L17P variation translates into protein S deficiency. Protein S-L17P causes its cytosolic mislocalization resulting in its proteasome-dependent degradation.
Topics: Animals; COS Cells; Chlorocebus aethiops; HEK293 Cells; Humans; Hydrophobic and Hydrophilic Interactions; Proteasome Endopeptidase Complex; Protein S; Protein Sorting Signals
PubMed: 34968852
DOI: 10.1016/j.thromres.2021.12.014 -
Food Chemistry Feb 2020This research aimed to explore the role of protein S-nitrosylation in regulating the tenderness of postmortem beef, from the perspective of μ-calpain autolysis and...
This research aimed to explore the role of protein S-nitrosylation in regulating the tenderness of postmortem beef, from the perspective of μ-calpain autolysis and protein proteolysis. Five bovine semimembranosus muscles were incubated with three treatments including S-nitrosoglutathione (GSNO, nitric oxide donor), normal saline and Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME, nitric oxide synthase inhibitor). The results showed that the level of protein S-nitrosylation was improved by GSNO treatment and reduced by L-NAME treatment (p < 0.05). Compared to the control, GSNO treatment had higher shear force while L-NAME treatment presented lower shear force at 7 d postmortem (p < 0.05). In addition, μ-calpain autolysis, myofibrillar protein and desmin degradation were reduced by GSNO treatment and accelerated by L-NAME treatment (p < 0.05). Therefore, it can be speculated that protein S-nitrosylation could affect beef tenderization by regulating the autolysis of μ-calpain and the degradation of myofibrillar proteins.
Topics: Animals; Arginine; Calpain; Cattle; Desmin; Protein S; Proteolysis; Red Meat
PubMed: 31622832
DOI: 10.1016/j.foodchem.2019.125616