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Applied and Environmental Microbiology Nov 2022Proteins immobilized on biosilica which have superior reactivity and specificity and are innocuous to natural environments could be useful biological materials in...
Proteins immobilized on biosilica which have superior reactivity and specificity and are innocuous to natural environments could be useful biological materials in industrial processes. One recently developed technique, living diatom silica immobilization (LiDSI), has made it possible to immobilize proteins, including multimeric and redox enzymes, via a cellular excretion system onto the silica frustule of the marine diatom . However, the number of application examples so far is limited, and the type of proteins appropriate for the technique is still enigmatic. Here, we applied LiDSI to six industrially relevant polypeptides, including protamine, metallothionein, phosphotriesterase, choline oxidase, laccase, and polyamine synthase. Protamine and metallothionein were successfully immobilized on the frustule as protein fusions with green fluorescent protein (GFP) at the N terminus, indicating that LiDSI can be used for polypeptides which are rich in arginine and cysteine. In contrast, we obtained mutants for the latter four enzymes in forms without green fluorescent protein. Immobilized phosphotriesterase, choline oxidase, and laccase showed enzyme activities even after the purification of frustule in the presence of 1% (wt/vol) octylphenoxy poly(ethyleneoxy)ethanol. An immobilized branched-chain polyamine synthase changed the intracellular polyamine composition and silica nanomorphology. These results illustrate the possibility of LiDSI for industrial applications. Proteins immobilized on biosilica which have superior reactivity and specificity and are innocuous to natural environments could be useful biological materials in industrial processes. Living diatom silica immobilization (LiDSI) is a recently developed technique for protein immobilization on the diatom frustule. We aimed to explore the possibility of using LiDSI for industrial applications by successfully immobilizing six polypeptides: (i) protamine (Oncorhynchus keta), a stable antibacterial agent; (ii) metallothionein (Saccharomyces cerevisiae), a metal adsorption molecule useful for bioremediation; (iii) phosphotriesterase (Sulfolobus solfataricus), a scavenger for toxic organic phosphates; (iv) choline oxidase (Arthrobacter globiformis), an enhancer for photosynthetic activity and yield of plants; (v) laccase (Bacillus subtilis), a phenol oxidase utilized for delignification of lignocellulosic materials; and (vi) branched-chain polyamine synthase (Thermococcus kodakarensis), which produces branched-chain polyamines important for DNA and RNA stabilization at high temperatures. This study provides new insights into the field of applied biological materials.
Topics: Diatoms; Green Fluorescent Proteins; Laccase; Silicon Dioxide; Peptides; Polyamines; Phosphoric Triester Hydrolases; Metallothionein; Protamines
PubMed: 36226967
DOI: 10.1128/aem.01153-22 -
Molecular and Cellular Endocrinology Dec 2020The spermatozoon is a highly differentiated cell with unique characteristics: it is mobile, thanks to its flagellum, and is very compact. The sperm cytoplasm is... (Review)
Review
The spermatozoon is a highly differentiated cell with unique characteristics: it is mobile, thanks to its flagellum, and is very compact. The sperm cytoplasm is extremely reduced, containing no ribosomes, and therefore does not allow translation, and its nucleus contains very closed chromatin, preventing transcription. This DNA compaction is linked to the loss of nucleosomes and the replacement of histones by protamines. Based on these characteristics, sperm was considered to simply deliver paternal DNA to the oocyte. However, some parts of the sperm DNA remain organized in a nucleosomal format, and bear epigenetic information. In addition, the nucleus and the cytoplasm contain a multitude of RNAs of different types, including non-coding RNAs (ncRNAs) which also carry epigenetic information. For a long time, these RNAs were considered residues of spermatogenesis. After briefly describing the mechanisms of compaction of sperm DNA, we focus this review on the origin and function of the different ncRNAs. We present studies demonstrating the importance of these RNAs in embryonic development and transgenerational adaptation to stress. We also look at other epigenetic marks, such as DNA methylation or post-translational modifications of histones, and show that they are sensitive to environmental stress and transmissible to offspring. The post-fertilization role of certain sperm-borne proteins is also discussed.
Topics: Animals; DNA; Embryonic Development; Epigenesis, Genetic; Female; Fertilization; Humans; Male; Mammals; Paternal Inheritance; Pregnancy; Protein Processing, Post-Translational; Spermatozoa; Stress, Physiological
PubMed: 32738444
DOI: 10.1016/j.mce.2020.110964 -
Urology Journal Jul 2020Recreational use of illicit drugs is one of the main factors affecting male fertility. However, the mechanisms of heroin smoke-associated damage to mature spermatozoa...
PURPOSE
Recreational use of illicit drugs is one of the main factors affecting male fertility. However, the mechanisms of heroin smoke-associated damage to mature spermatozoa are still completely unknown. The aim of this study was to concomitantly examine the levels of protamine-2 gene and protein concentrations, the amount of miRNA-122 in seminal plasma and semen analysis findings in heroin-addicted men.
MATERIALS AND METHODS
In a case control study, twenty-four fertile men that lacked any recreational drug abuse were considered as the healthy group, and 24 addicted men who used only heroin for at least four months were selected as the addicted group. Semen samples were gathered by masturbation after 2 - 5 days of sexual abstinence. Following the preparation of a semen analysis by computer-assisted sperm analysis according to WHO (2010), the level of protamine-2 gene expression in sperm and miRNA-122 in seminal plasma was measured using real-time sqPCR. Also, protamine-2 protein concentrations were quantified by nuclear protein extraction, SDS-Page and western blotting.
RESULTS
Among the studied variables, body mass index (27.75±0.88 vs. 22.30±0.36, p=0.001), seminal pH (7.79±0.06 vs. 7.58±0.06, p=0.003), white blood cell count in semen (1.69±0.41 vs. 8.61±1.73, p=0.001), motility (65.51±2.57 vs. 41.96±3.58, p=0.001) and survival rate (87.41±1.00 vs. 71.50±4.59, p=0.002) of sperm cells was significantly different between the healthy and addicted groups. In addition, the levels of protamine-2 gene and protein expression in the addicted group (0.05±0.02 and 0.10±0.02, respectively) were significantly lower than the healthy group (3.59±0.94 and 0.27±0.06, respectively) (p=0.002 and p=0.017, respectively). Seminal miRNA-122 levels in addicted men (3.51±0.73) were statistically higher than in healthy men (1.52±0.54) (p=0.034). However, there were some significant relationship between the studied parameters and addiction (p<0.05).
CONCLUSION
This is one study on human infertility that evaluates the effects of heroin on protamine deficiency and seminal small RNAs expression levels. Heroin abuse may lead to male infertility by causing leukocytospermia, asthenozoospermia, protamine deficiency, and seminal plasma miRNA profile alteration.
Topics: Adult; Case-Control Studies; Correlation of Data; Heroin Dependence; Humans; Male; MicroRNAs; Protamines; Semen; Semen Analysis; Spermatozoa
PubMed: 32748386
DOI: 10.22037/uj.v16i7.5747 -
Materials (Basel, Switzerland) Sep 2019Protamine is an antimicrobial peptide extracted from fish. In this study, we loaded protamine onto dicalcium phosphate anhydride (DCPA), a dental material. Protamine was...
Protamine is an antimicrobial peptide extracted from fish. In this study, we loaded protamine onto dicalcium phosphate anhydride (DCPA), a dental material. Protamine was loaded by stirring DCPA into a protamine solution. To explore the antimicrobial activity of the materials, we cultivated on fabricated discs for 24 h. When was cultivated on the discs under no sucrose conditions, the loaded protamine was not released, and the ratio of dead bacteria increased on the surface of P (125) DCPA (half of the saturated level of protamine (125 ppm protamine) was loaded). Aside from P (500) DCPA (saturated level of protamine was loaded), some protamine was released, and the number of planktonic bacteria in the supernatant decreased. Using medium containing 1% sucrose, the release of protamine was promoted from P (125) DCPA due to lowered pH. However, lowering of the pH decreased the antimicrobial activity of protamine. On the other hand, P (500) DCPA released protamine before the pH was lowered, and biofilm formation was inhibited. The loaded protamine expressed antimicrobial activity, both on the surface of the materials and in the surrounding environment. The interaction of loaded protamine with calcium phosphates could promote the application of protamine in the dental field.
PubMed: 31480654
DOI: 10.3390/ma12172816 -
Journal of Environmental Science and... 2023This study was conducted to assess the impact of hubble-bubble smoking on global DNA methylation, DNA fragmentation; protamine deficiency of spermatozoa, and to...
This study was conducted to assess the impact of hubble-bubble smoking on global DNA methylation, DNA fragmentation; protamine deficiency of spermatozoa, and to determine whether the transcription levels of the and genes are different in hubble-bubble smokers compared to nonsmokers. Five hundred semen samples were collected from males with an average age of 32.2 ± 6.1 years (300 hubble-bubble smokers "60%" and 200 nonsmokers "40%"). The nucleic acid was isolated from purified sperm, then ELISA and qPCR were used to evaluate the global DNA methylation and transcription level of and , respectively. A significant elevation in global DNA methylation, protamine deficiency, and DNA fragmentation was found in hubble-bubble smokers compared to nonsmokers (P < 0.0001). A significant decline was shown in transcription levels of and genes in hubble-bubble compared to nonsmokers (P < 0.0001). Additionally, a down-regulation in the transcription levels of and was revealed in hubble-bubble compared to nonsmokers with fold change (0.0001 and 0.007, respectively). In conclusion, this study provided proof that hubble-bubble smoking has a negative impact on global DNA methylation, DNA fragmentation, protamine deficiency, and the transcription of and genes in spermatozoa, and these findings influence negatively males' fecundity.
Topics: Humans; Male; Adult; Histones; DNA Methylation; Semen; Protamines; Spermatozoa; Smoking; Infertility, Male
PubMed: 36744325
DOI: 10.1080/10934529.2023.2174326 -
Evolution; International Journal of... Aug 2021Post-copulatory sexual selection is thought to influence the evolution of genes involved in reproduction. However, the detection of straightforward effects has been...
Post-copulatory sexual selection is thought to influence the evolution of genes involved in reproduction. However, the detection of straightforward effects has been proven difficult due to the complexity and diversity of reproductive landscapes found in different taxa. Here, we compare the possible effect of relative testes mass as a sperm competition proxy on protamine genotype (protamine 1/protamine 2 ratio) and the link to sperm head phenotype in two rodent groups, mice, and voles. In mice, protamine expression ratios were found to increase from low values toward a 1:1 ratio in a positive association with testes mass, and relative sperm head area. In contrast, in voles, decreasing protamine expression ratios were found in species with larger testes but, surprisingly, they range from high values, again toward a 1:1 ratio, and showing a negative correlation with relative sperm head area. Altogether, we found differences in the way protamines seem to be selected and involved in adaptations of the sperm head in voles and mice. However, sexual selection driven by sperm competition seems to exhibit a common evolutionary pattern in both groups toward an equilibrium in the expression of the two protamines.
Topics: Animals; Male; Mice; Protamines; Rodentia; Sexual Selection; Sperm Head; Spermatozoa
PubMed: 34224143
DOI: 10.1111/evo.14305 -
ACS Sensors Oct 2022The last decade has witnessed a rapid development of nano- and microparticle-based optical ion sensors, including ion-selective optodes (ISOs). While the application of...
The last decade has witnessed a rapid development of nano- and microparticle-based optical ion sensors, including ion-selective optodes (ISOs). While the application of nano-ISOs has shown promising performance for sensing inorganic ions, polyion sensing using nanoscale ISOs has encountered significant interference in complex samples such as blood plasma. Recently, we have reported on a new polyion sensing principle that operates through a novel mechanism to overcome this challenge. The new sensing mechanism showed improved characteristics not observed with conventional ion-exchange type sensors, but the precise mechanism of operation remained thus far unclear. This paper aims to clarify how protamine, the arginine-rich target polycation, behaves during optical signal transduction to give dramatically improved selectivity. Based on thermodynamic data, sensor performance and ζ-potential analysis, two discrete phases of protamine extraction are identified. Initially, protamine extracts into the bulk nanosensor phase, a process that is concurrent with the optical signal change. This is then followed by protamine accumulation onto the nanosensor surface, which starts only upon saturation of the optical signal change. The data indicate that the improved selectivity is due to the inability of small ions to form a sufficiently strong interaction with an active sensing ingredient, DNNS. Any exchange of one inorganic cation for another therefore remains optically silent, suppressing matrix effects. Moreover, the recognition of protamine is shown to be an exhaustive extraction process, making the response independent of the nature and concentration of the initial small cation in the nanosensor phase.
Topics: Protamines; Cations
PubMed: 36121929
DOI: 10.1021/acssensors.2c01599 -
Frontiers in Endocrinology 2022Male fertility throughout life hinges on the successful production of motile sperm, a developmental process that involves three coordinated transitions: mitosis,... (Review)
Review
Male fertility throughout life hinges on the successful production of motile sperm, a developmental process that involves three coordinated transitions: mitosis, meiosis, and spermiogenesis. Germ cells undergo both mitosis and meiosis to generate haploid round spermatids, in which histones bound to the male genome are replaced with small nuclear proteins known as protamines. During this transformation, the chromatin undergoes extensive remodeling to become highly compacted in the sperm head. Despite its central role in spermiogenesis and fertility, we lack a comprehensive understanding of the molecular mechanisms underlying the remodeling process, including which remodelers/chaperones are involved, and whether intermediate chromatin proteins function as discrete steps, or unite simultaneously to drive successful exchange. Furthermore, it remains largely unknown whether more nuanced interactions instructed by protamine post-translational modifications affect chromatin dynamics or gene expression in the early embryo. Here, we bring together past and more recent work to explore these topics and suggest future studies that will elevate our understanding of the molecular basis of the histone-to-protamine exchange and the underlying etiology of idiopathic male infertility.
Topics: Chromatin; Histones; Humans; Male; Protamines; Semen; Spermatozoa
PubMed: 35813619
DOI: 10.3389/fendo.2022.895502 -
Cancers Apr 2022In maturing sperm cells, a major genome re-organization takes place, which includes a global increase in the acetylation of histones prior to their replacement by... (Review)
Review
In maturing sperm cells, a major genome re-organization takes place, which includes a global increase in the acetylation of histones prior to their replacement by protamines, the latter being responsible for the tight packaging of the male genome. Understanding the function of the oncogenic BRD4-NUT fusion protein in NUT carcinoma (NC) cells has proven to be essential in uncovering the mechanisms underlying histone hyperacetylation in spermatogenic cells. Indeed, these studies have revealed the mechanism by which a cooperation between BRD4, a bromodomain factor of the BET family, NUT, a normally testis-specific factor, and the histone acetyltransferase p300, induces the generation of hyperacetylated chromatin domains which are present in NC cells. The generation of ko mice enabled us to demonstrate a genetic interaction between and , encoding BRDT, a testis-specific BRD4-like factor. Indeed, in spermatogenic cells, NUT and p300 interact, which results in an increased acetylation of histone H4 at both positions K5 and K8. These two positions, when both acetylated, are specifically recognized by the first bromodomain of BRDT, which then mediates the removal of histone and their replacement by protamines. Taken together, these investigations show that the fusion of NUT to BRD4 in NUT Carcinoma cells reconstitutes, in somatic cells, a functional loop, which normally drives histone hyperacetylation and chromatin binding by a BET factor in spermatogenic cells.
PubMed: 35565363
DOI: 10.3390/cancers14092234 -
Food Microbiology Apr 2022Hepatitis A virus (HAV) continues to be a public health concern and has caused large foodborne outbreaks and economic losses worldwide. Rapid detection of HAV in foods...
Efficient capturing and sensitive detection of hepatitis A virus from solid foods (green onion, strawberry, and mussel) using protamine-coated iron oxide (FeO) magnetic nanoparticles and real-time RT-PCR.
Hepatitis A virus (HAV) continues to be a public health concern and has caused large foodborne outbreaks and economic losses worldwide. Rapid detection of HAV in foods can help to confirm the source of outbreaks in a timely manner and prevent more people getting infected. In order to efficiently detect HAV at low levels of contamination in foods, rapid and easy-to-use techniques are required to separate and concentrate viral particles to a small volume. In the current study, HAV particles were eluted from green onion, strawberry, and mussel using glycine buffer (0.05 M glycine, 0.14 M NaCl, 0.2% (v/v) Tween 20, pH 9.0) and suspended viral particles were captured using protamine-coated magnetic nanoparticles (PMNPs). This process caused a selective concentration of the viral particles, which could be followed by quantitative real-time RT-PCR analysis. Results showed that pH, NaCl concentration, and PMNP amount used for the capturing had significant effects on the recovery efficiency of HAV (P < 0.05). The highest recovery rate was obtained at pH 9.0, 0.14 M NaCl, and 50 μL of PMNPs. The optimized PMNP capturing method enabled the rapid capture and concentration of HAV. A sensitive real-time RT-PCR test was developed with detection limits of 8.3 × 10 PFU/15 g, 8.3 × 10 PFU/50 g, and 8.3 × 10 PFU/5 g of HAV in green onion, strawberry, and mussel, respectively. In conclusion, the PMNP method is rapid and convenient in capturing HAV from complex solid food samples and can generate concentrated HAV sample solutions suitable for high-sensitivity real time RT-PCR detection of the virus.
Topics: Animals; Bivalvia; Ferric Compounds; Food Contamination; Fragaria; Hepatitis A virus; Magnetite Nanoparticles; Onions; Protamines; RNA, Viral; Reverse Transcriptase Polymerase Chain Reaction
PubMed: 34809947
DOI: 10.1016/j.fm.2021.103921