scholarly journals A novel posttranslational modification of histone, H3 S-sulfhydration, is down-regulated in asthenozoospermic sperm

2021 ◽  
Author(s):  
Qi Qi ◽  
Hongjie Pan ◽  
Ning Jiang ◽  
Meixin Zhang ◽  
Shenfei Sun ◽  
...  
Keyword(s):  
Male Infertility ◽  
Posttranslational Modification ◽  
Protein S ◽  
Human Sperm ◽  
Spermatogonial Stem Cell ◽  
Stem Cell Line ◽  
Sperm Protein ◽  
Progressive Motility ◽  
Biotin Switch ◽  
First Time

AbstractOxidative stress is one of the major causes leading to male infertility including asthenozoospermia. Hydrogen sulfide (H2S) has been widely recognized to be a potent antioxidant whose role is partially implemented by protein S-sulfhydration. However, protein S-sulfhydration has not been reported in germ cells. Therefore, we investigated whether asthenozoospermia could be associated with sperm protein S-sulfhydration. S-sulfhydrated proteins in human sperm were enriched via biotin-switch assay and analyzed using LC-MS/MS spectrometry. Two hundred forty-four S-sulfhydrated proteins were identified. Importantly, we validated that sperm histones H3.1 and H3.3 were the S-sulfhydrated proteins. Their S-sulfhydrated amino acid residue was Cysteine111. Abundances of S-sulfhydrated H3 (sH3) and S-sulfhydrated H3.3 (sH3.3) were significantly down-regulated in asthenozoospermic sperm, compared with the fertile controls, and were significantly correlated with progressive motility. Retinoic acid (RA) up-regulated level of sH3.3 in primary round spermatids and the C18-4 cells (a mouse spermatogonial stem cell line). Overexpression of the mutant H3.3 (Cysteine111 was replaced with serine) affected expression of 759 genes and raised growth rate of C18-4 cells. For the first time, S-sulfhydration H3 and H3.3 were demonstrated in the present study. Our results highlight that aberrant S-sulfhydration of H3 is a new pathophysiological basis in male infertility.

scholarly journals Expression of PPP3CC and not PPP3R2 is associated with asthenozoospermia

2021 ◽  
Keyword(s):  
Male Infertility ◽  
Protein Expression ◽  
Protein Phosphatase ◽  
Calcineurin Inhibitors ◽  
Human Sperm ◽  
Regulatory Subunit ◽  
Regulatory Subunits ◽  
Protein Levels ◽  
Progressive Motility ◽  
Mrna And Protein Expression

Background and objective: Protein phosphatase 3 catalytic subunit gamma (PPP3CC) and protein phosphatase 3 regulatory subunit B, beta (PPP3R2) are respectively the catalytic and regulatory subunits of calcineurin in sperm. Deficiency in either protein causes impaired sperm motility leading to male infertility. Many cases of sterility are attributed to asthenozoospermia (AZS); however, it remains unknown whether PPP3CC and PPP3R2 are related to AZS. Material and methods: Quantitative PCR and Western blotting were used to investigate the expression levels of PPP3CC and PPP3R2 in the spermatozoa of patients with AZS and to explore the clinical significance. Results: Two calcineurin inhibitors cyclosporine A (CsA) and tacrolimus (FK506) markedly impaired the total motility and progressive motility of human sperm, indicating that PPP3CC or PPP3R2 might be involved in AZS. PPP3CC mRNA and protein expression was lower in the ejaculated spermatozoa of patients with AZS than in normal sperm (NS). Correlation analysis showed that PPP3CC protein expression correlated positively with progressive motility (r = 0.2592, P < 0.05); however, there were no significant differences in PPP3R2 mRNA and protein levels between AZS and NS. Conclusion: These findings suggest that the abnormal expression of PPP3CC rather than PPP3R2 might be a pathological factor or indicator in AZS. Thus, PPP3CC may be a potential therapeutic or diagnostic target for some cases of male infertility.

Human sperm superoxide anion generation and correlation with semen quality in patients with male infertility

2004 ◽  
Vol 82 (4) ◽  
pp. 871-877 ◽  
Author(s):  
T SAID ◽  
A AGARWAL ◽  
R SHARMA ◽  
E MASCHA ◽  
S SIKKA ◽  
...  
Keyword(s):  
Male Infertility ◽  
Superoxide Anion ◽  
Semen Quality ◽  
Human Sperm ◽  
Superoxide Anion Generation

Abstract 484: Nitrosative Stress Enhances S-nitrosation Of Peroxiredoxins In The Heart

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Michael Reinartz ◽  
Zhaoping Ding ◽  
Axel Gödecke ◽  
Jürgen Schrader
Keyword(s):  
Nitric Oxide ◽  
Nitrosative Stress ◽  
Protein S ◽  
List Type ◽  
Basal Expression ◽  
Protein Thiols ◽  
Wide Range ◽  
Well Data ◽  
Oxide Synthase ◽  
Biotin Switch

Nitric oxide (NO) is produced by different isoforms of NO-synthases and operates as a mediator of important cell signaling pathways. To explore thiol-based protein modifications in a situation of nitrosative stress, two transgenic mouse models recently generated in our laboratory were used: Cardiac specific overexpression of inducible nitric oxide synthase (iNOS) (tg-iNOS + ) and tg-iNOS + with concomitant myoglobin-deficiency (tg-iNOS + /myo −/− ). Protein S-nitrosation, an important redox-based posttranslational modification, revealed no differences between WT and tg-iNOS + hearts as measured by the biotin-switch assay and 2-D PAGE. Even in the absence of myoglobin - an efficient endogenous NO-oxidase - the protein S-nitrosation pattern for nearly all the detected proteins (>40) remained unchanged in the tg-iNOS + /myo −/− hearts, with the exception of three proteins. Tandem mass spectrometry uncovered these proteins as peroxiredoxins (Prx II, III, VI), which are known to possess peroxidase activity, whereby hydrogen peroxide, peroxynitrite and a wide range of organic hydroperoxides are reduced and detoxified. To prove whether the higher abundance of the Prxs was due to enhanced S-nitrosation or due to changes in their basal expression levels, immunoblotting with specific antibodies was applied and revealed upregulation of Prx VI in tg-iNOS + /myo −/− hearts. The other proteins found to be S-nitrosated were identified as well. Data mining indicated a significant overlap of these proteins with proteins becoming glutathiolated. Protein glutathiolation detected by immunoblotting was enhanced in the tg-iNOS + hearts and even more so in the tg-iNOS + /myo −/− hearts. We conclude that protein glutathiolation in our transgenic model of nitrosative stress is important to protect protein thiols from irreversible oxidation. The upregulation of antioxidant proteins like Prx VI appears to be an additional mechanism to antagonize an excess of reactive oxygen/nitrogen species. Enhanced S-nitrosation of the Prxs may serve a new function in the signalling cascade coping with nitrosative stress.

S-nitrosylation of CYR61 protein reduces triple-negative breast cancer metastasis ability

2020 ◽  
Author(s):  
Lee Jia ◽  
Yusheng Lu ◽  
Sudan He ◽  
Huanzhang Xie ◽  
Chunlian Zhong ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Mass Spectrometry ◽  
Intravenous Injection ◽  
Triple Negative Breast Cancer ◽  
Cancer Metastasis ◽  
Triple Negative ◽  
3D Structure ◽  
Protein S ◽  
Adhesion Assay ◽  
Biotin Switch

Abstract BackgroundTriple-negative breast cancer (TNBC) is the most difficult cancer to be treated. TNBC expresses high level of matricellular cysteine-rich protein CYR61/CCN1 that plays a key role in producing cancer metastases and is an important target for metastasis chemoprevention. Nitric oxide (NO) can covalently bind to the thiol group of cysteines (termed S-nitrosylation) resulting in regulation of the targeted protein functions. MethodsProtein S-nitrosylation were detected by biotin-switch assay and western blotting assay. CYR61 protein S-nitrosylated sites and 3D structure were determined by mass spectrometry and MODELLER software. Adhesion assay, cell morphology assay, wound healing assay and transwell invasion assay were used to evaluate effects of CYR61 S-nitrosylation on the cell metastatic ability. In vivo metastasis activity of CYR61 S-nitrosylation were tested by intravenous injection and mammary xenograft implantation mouse metastatic models.ResultsS-nitrosylation by GSNO of CYR61 reached a plateau quickly and was confirmed by spectroscopic analysis and biotin-switch assay. Mass-spectrometry proteomic analysis revealed that S-nitrosylation predominantly occurred at Cys100, Cys117, Cys229 and Cys239, resulting in CYR61 structure relaxed and unstable evidenced by protein structure modeling. S-nitrosylation of MDA-MB-231 cells, their CYR61-overexpressed and CYR61–silenced counterparts significantly attenuated the metastatic ability of these cells, including their ability of adhesion, mobility, invasion, and interplay with platelets, and made the adhered cells unattached. The attenuation in metastatic ability proportionally increased with the degree of S-nitrosylation to CYR61 naturally-expressed or genetically-manipulated cells, and was demonstrated in mice, where, S-nitrosylation of these cell lines not only inhibited their acute seeding to lungs after an intravenous injection, but also inhibited the late development of these cells into the metastatic nodes after mammary xenograft implantation. Furthermore, orthotopically-implanted MDA-MB-231 developed mammary tumors and later lung metastasis; whereas, the same cells with S-nitrosylation developed no tumor and metastasis at all. Conclusionwe present the first evidence that S-nitrosylation of CYR61 can significantly inhibit metastatic aggressiveness of the TNBC MDA-MB-231 cells. This conceptual creative study opens a new avenue to prevent the most aggressive TNBC from metastases by S-nitrosylation to CYR61.

Expression of human sperm protein 17 in melanophages of cutaneous melanocytic lesions

2004 ◽  
Vol 150 (4) ◽  
pp. 780-782 ◽  
Author(s):  
B. Franceschini ◽  
F. Grizzi ◽  
P. Colombo ◽  
G. Soda ◽  
K. Bumm ◽  
...  
Keyword(s):  
Human Sperm ◽  
Sperm Protein ◽  
Melanocytic Lesions ◽  
Sperm Protein 17

Transient scrotal hyperthermia affects human sperm DNA integrity, sperm apoptosis, and sperm protein expression

Andrology ◽  
2016 ◽  
Vol 4 (6) ◽  
pp. 1054-1063 ◽  
Author(s):  
M. Rao ◽  
W. Xia ◽  
J. Yang ◽  
L.-X. Hu ◽  
S.-F. Hu ◽  
...  
Keyword(s):  
Protein Expression ◽  
Human Sperm ◽  
Dna Integrity ◽  
Sperm Protein ◽  
Sperm Dna Integrity ◽  
Sperm Dna

scholarly journals Impact of Oxidative Stress and Protein S-Glutathionylation in Aortic Valve Sclerosis Patients with Overt Atherosclerosis

2019 ◽  
Vol 8 (4) ◽  
pp. 552 ◽  
Author(s):  
Vincenza Valerio ◽  
Veronika A. Myasoedova ◽  
Donato Moschetta ◽  
Benedetta Porro ◽  
Gianluca L. Perrucci ◽  
...  
Keyword(s):  
Aortic Valve ◽  
Smooth Muscle Actin ◽  
Artery Bypass ◽  
Protein S ◽  
Aortic Valve Sclerosis ◽  
Cell Adhesion Molecule 1 ◽  
First Time ◽  
Glutathione Homeostasis

Aortic valve sclerosis (AVSc) is characterized by non-uniform thickening of the leaflets without hemodynamic changes. Endothelial dysfunction, also caused by dysregulation of glutathione homeostasis expressed as ratio between its reduced (GSH) and its oxidised form (GSSG), could represent one of the pathogenic triggers of AVSc. We prospectively enrolled 58 patients with overt atherosclerosis and requiring coronary artery bypass grafting (CABG). The incidence of AVSc in the studied population was 50%. The two groups (No-AVSc and AVSc) had similar clinical characteristics. Pre-operatively, AVSc group showed significantly lower GSH/GSSG ratio than No-AVSc group (p = 0.02). Asymmetric dimethylarginine (ADMA) concentration was significantly higher in AVSc patients compared to No-AVSc patients (p < 0.0001). Explanted sclerotic aortic valves presented a significantly increased protein glutathionylation (Pr-SSG) than No-AVSc ones (p = 0.01). In vitro, inhibition of glutathione reductase caused β-actin glutathionylation, activation of histone 2AX, upregulation of α2 smooth muscle actin (ACTA2), downregulation of platelet and endothelial cell adhesion molecule 1 (PECAM1) and cadherin 5 (CDH5). In this study, we showed for the first time that the dysregulation of glutathione homeostasis is associated with AVSc. We found that Pr-SSG is increased in AVSc leaflets and it could lead to EndMT via DNA damage. Further studies are warranted to elucidate the causal role of Pr-SSG in aortic valve degeneration.

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