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The Journal of Neuroscience : the... Jul 2022Synaptotagmin-1 (Syt1) is a vesicular calcium sensor required for synchronous neurotransmitter release, composed of a single-pass transmembrane domain linked to two C2...
Synaptotagmin-1 (Syt1) is a vesicular calcium sensor required for synchronous neurotransmitter release, composed of a single-pass transmembrane domain linked to two C2 domains (C2A and C2B) that bind calcium, acidic lipids, and SNARE proteins that drive fusion of the synaptic vesicle with the plasma membrane. Despite its essential role, how Syt1 couples calcium entry to synchronous release is poorly understood. Calcium binding to C2B is critical for synchronous release, and C2B additionally binds the SNARE complex. The C2A domain is also required for Syt1 function, but it is not clear why. Here, we asked what critical feature of C2A may be responsible for its functional role and compared this to the analogous feature in C2B. We focused on highly conserved poly-lysine patches located on the sides of C2A (K189-192) and C2B (K324-327). We tested effects of charge-neutralization mutations in either region (Syt1 and Syt1) side by side to determine their relative contributions to Syt1 function in cultured cortical neurons from mice of either sex and in single-molecule experiments. Combining electrophysiological recordings and optical tweezers measurements to probe dynamic single C2 domain-membrane interactions, we show that both C2A and C2B polybasic patches contribute to membrane binding, and both are required for evoked release. The size of the readily releasable vesicle pool and the rate of spontaneous release were unaffected, so both patches are likely required specifically for synchronization of release. We suggest these patches contribute to cooperative membrane binding, increasing the overall affinity of Syt1 for negatively charged membranes and facilitating evoked release. Synaptotagmin-1 is a vesicular calcium sensor required for synchronous neurotransmitter release. Its tandem cytosolic C2 domains (C2A and C2B) bind calcium, acidic lipids, and SNARE proteins that drive fusion of the synaptic vesicle with the plasma membrane. How calcium binding to Synaptotagmin-1 leads to release and the relative contributions of the C2 domains are unclear. Combining electrophysiological recordings from cultured neurons and optical tweezers measurements of single C2 domain-membrane interactions, we show that conserved polybasic regions in both domains contribute to membrane binding cooperatively, and both are required for evoked release, likely by increasing the overall affinity of Synaptotagmin-1 for acidic membranes.
Topics: Animals; C2 Domains; Calcium; Lipids; Mice; Neurotransmitter Agents; SNARE Proteins; Synaptotagmin I
PubMed: 35701163
DOI: 10.1523/JNEUROSCI.1385-21.2022 -
Drug Delivery Nov 2020A novel β-cyclodextrin pendant polymer (ε-PL-CD), composed of poly(ε-lysine) (ε-PL) main chain and glycine-β-cyclodextrin (Gly-CD) side chains, was prepared by a...
A novel β-cyclodextrin pendant polymer (ε-PL-CD), composed of poly(ε-lysine) (ε-PL) main chain and glycine-β-cyclodextrin (Gly-CD) side chains, was prepared by a simple two-step procedure. The ε-PL-CD was investigated as a drug carrier of hydrophobic drug scutellarin (SCU). The characterization and complexation mode of the SCU:ε-PL-CD were researched in both solution and solid state by means of photoluminescence spectroscopy, H and 2D NMR, X-Ray powder diffraction (XRPD), thermal gravimetric analysis, Particle size and Zeta potential. The solubility test indicated that the solubilizing ability of SCU:ε-PL-CD was significantly improved compared with SCU:β-CD and free SCU. Besides, in cell experiment, it was found that SCU:ε-PL-CD has a strong inhibitory effect on the growth and invasion of tumor cells. The present study provides useful information for ε-PL-CD as a drug carrier material.
Topics: Apigenin; Cellulose; Crystallography, X-Ray; Cyclodextrins; Drug Carriers; Drug Delivery Systems; Glucuronates; Humans; Magnetic Resonance Spectroscopy; Molecular Structure; Nanoparticles; Particle Size; Polylysine; Solubility
PubMed: 33307844
DOI: 10.1080/10717544.2020.1856223 -
Foods (Basel, Switzerland) Jul 2023In the present study, electrospun nanofiber mats were fabricated by mixing different ratios (96:4, 95:5, 94:6, 93:7, and 92:8) of Persian gum (PG) and poly (ethylene...
Fabrication of ɛ-Polylysine-Loaded Electrospun Nanofiber Mats from Persian Gum-Poly (Ethylene Oxide) and Evaluation of Their Physicochemical and Antimicrobial Properties.
In the present study, electrospun nanofiber mats were fabricated by mixing different ratios (96:4, 95:5, 94:6, 93:7, and 92:8) of Persian gum (PG) and poly (ethylene oxide) (PEO). The SEM micrographs revealed that the nanofibers obtained from 93% PG and 7% PEO were bead-free and uniform. Therefore, it was selected as the optimized ratio of PG:PEO for the development of antimicrobial nanofibers loaded with ɛ-Polylysine (ɛ-PL). All of the spinning solutions showed pseudoplastic behavior and the viscosity decreased by increasing the shear rate. Additionally, the apparent viscosity, G', and G″ of the spinning solutions increased as a function of PEO concentration, and the incorporation of ɛ-PL did not affect these parameters. The electrical conductivity of the solutions decreased when increasing the PEO ratio and with the incorporation of ɛ-PL. The X-ray diffraction (XRD) and Fourier-transform infrared (FTIR) spectra showed the compatibility of polymers. The antimicrobial activity of nanofibers against () and () was investigated, and the samples loaded with ɛ-PL demonstrated stronger antimicrobial activity against
PubMed: 37444326
DOI: 10.3390/foods12132588 -
Journal of Food Protection Jul 2024Salmonella is capable of surviving dehydration within various foods, such as dried fruit. Dried fruit, including apple slices, have been the subject of product recalls...
Salmonella is capable of surviving dehydration within various foods, such as dried fruit. Dried fruit, including apple slices, have been the subject of product recalls due to contamination with Salmonella. A study was conducted to determine the fate of Salmonella on apple slices, following immersion in three antimicrobial solutions (viz., ε-polylysine [epsilon-polylysine or EP], sodium bisulfate [SBS], or peracetic acid [PAA]), and subsequent hot air dehydration. Gala apples were aseptically cored and sliced into 0.4 cm thick rings, bisected, and inoculated with a five-strain composite of desiccation-resistant Salmonella, to a population of 8.28 log CFU/slice. Slices were then immersed for 2 min in various concentrations of antimicrobial solutions, including EP (0.005, 0.02, 0.05, and 0.1%), SBS (0.05, 0.1, 0.2, and 0.3%), PAA (18 or 42 ppm), or varying concentrations of PAA + EP, and then dehydrated at 60°C for 5 h. Salmonella populations in positive control samples (inoculated apple slices washed in sterile water) declined by 2.64 log after drying. In the present study, the inactivation of Salmonella, following EP and SBS treatments, increased with increasing concentrations, with maximum reductions of 3.87 and 6.20 log (with 0.1 and 0.3% of the two compounds, respectively). Based on preliminary studies, EP concentrations greater than 0.1% did not result in lower populations of Salmonella. Pretreatment washes with either 18 or 42 ppm of PAA inactivated Salmonella populations by 4.62 and 5.63 log, respectively, following desiccation. Combining PAA with up to 0.1% EP induced no greater population reductions of Salmonella than washing with PAA alone. The addition of EP to PAA solutions appeared to destabilize PAA concentrations, reducing its biocidal efficacy. These results may provide antimicrobial predrying treatment alternatives to promote the reduction of Salmonella during commercial or consumer hot air drying of apple slices.
Topics: Malus; Peracetic Acid; Salmonella; Polylysine; Food Microbiology; Humans; Colony Count, Microbial; Sulfates; Food Preservation; Dose-Response Relationship, Drug; Desiccation; Food Contamination; Food Handling; Consumer Product Safety
PubMed: 38734414
DOI: 10.1016/j.jfp.2024.100297 -
Biosensors Sep 2022A label-free fipronil aptasensor was built based on Polylysine-black phosphorus nanosheets composition (PLL-BPNSs) and Au nanoparticles (AuNPs). A PLL-BP modified glassy...
A label-free fipronil aptasensor was built based on Polylysine-black phosphorus nanosheets composition (PLL-BPNSs) and Au nanoparticles (AuNPs). A PLL-BP modified glassy carbon electrode (GCE) was fabricated by combining BP NSs and PLL, which included a considerable quantity of -NH. Au nanoparticles (AuNPs) were placed onto the GCE, and PLL-BPNSs bonded to Au NPs firmly by assembling. The thiolated primers were then added and fixed using an S-Au bond, and competitive binding of the fipronil aptamer was utilized for fipronil quantitative assessment. The sensor's performance was evaluated using differential pulse voltammetry (DPV) method. The linear equation is ΔI (μA) = 13.04 logC + 22.35, while linear correlation coefficient R is 0.998, and detection limit is 74 pg/mL (0.17 nM) when the concentration of fipronil is 0.1 ng/mL-10 μg/mL. This aptasensor can apply to quantitative detection of fipronil.
Topics: Gold; Aptamers, Nucleotide; Biosensing Techniques; Metal Nanoparticles; Phosphorus; Polylysine; Electrodes; Carbon; Limit of Detection; Electrochemical Techniques
PubMed: 36290913
DOI: 10.3390/bios12100775 -
Pharmaceutics Jul 2021Despite being a mainstay of clinical cancer treatment, chemotherapy is limited by its severe side effects and inherent or acquired drug resistance. Nanotechnology-based...
Despite being a mainstay of clinical cancer treatment, chemotherapy is limited by its severe side effects and inherent or acquired drug resistance. Nanotechnology-based drug-delivery systems are widely expected to bring new hope for cancer therapy. These systems exploit the ability of nanomaterials to accumulate and deliver anticancer drugs at the tumor site via the enhanced permeability and retention effect. Here, we established a novel drug-delivery nanosystem based on amphiphilic peptide dendrimers (AmPDs) composed of a hydrophobic alkyl chain and a hydrophilic polylysine dendron with different generations (AmPD KK and AmPD KKK). These AmPDs assembled into nanoassemblies for efficient encapsulation of the anti-cancer drug doxorubicin (DOX). The AmPDs/DOX nanoformulations improved the intracellular uptake and accumulation of DOX in drug-resistant breast cancer cells and increased permeation in 3D multicellular tumor spheroids in comparison with free DOX. Thus, they exerted effective anticancer activity while circumventing drug resistance in 2D and 3D breast cancer models. Interestingly, AmPD KK bearing a smaller peptide dendron encapsulated DOX to form more stable nanoparticles than AmPD KKK bearing a larger peptide dendron, resulting in better cellular uptake, penetration, and anti-proliferative activity. This may be because AmPD KK maintains a better balance between hydrophobicity and hydrophilicity to achieve optimal self-assembly, thereby facilitating more stable drug encapsulation and efficient drug release. Together, our study provides a promising perspective on the design of the safe and efficient cancer drug-delivery nanosystems based on the self-assembling amphiphilic peptide dendrimer.
PubMed: 34371783
DOI: 10.3390/pharmaceutics13071092 -
Frontiers in Nutrition 2022The potential of bio-preservatives, namely, nisin, natamycin, and polylysine, as viable alternatives to chemical preservatives for storage of tender coconut water (TCW)...
The potential of bio-preservatives, namely, nisin, natamycin, and polylysine, as viable alternatives to chemical preservatives for storage of tender coconut water (TCW) during refrigerated storage (5 ± 2°C) was explored. Bio-preservative treatments were carried out after optimized heat treatment (85°C for 5 min) of TCW to establish its storage characteristics. Various concentrations (up to 125 ppm) of bio-preservatives were used for the preservation, and quality parameters of resultant TCW were assessed based on physicochemical characteristics and Food and Agriculture Organization (FAO) guidelines and statistical analysis applied. Analysis of variance (ANOVA) and test revealed that pH and overall acceptability (OA) are the major governing factors that determine spoilage of TCW ( < 0.05). Overall, the polylysine combination was found to be most effective in ensuring quality retention of TCW. It was concluded that pasteurized TCW shelf life could be extended up to 20 days using bio-preservatives.
PubMed: 36211480
DOI: 10.3389/fnut.2022.977655 -
Biophysical Journal Jul 2021The structure-driven assembly of multimeric protein complexes and the formation of intracellular phase-like protein condensates have been the subject of intense...
The structure-driven assembly of multimeric protein complexes and the formation of intracellular phase-like protein condensates have been the subject of intense research. However, the assembly of larger superstructures comprising cellular components, such as protein nanoparticles driven by general physical rather than specific biochemical interactions, remains relatively uncharacterized. Here, we use gas vesicles (GVs)-genetically encoded protein nanoparticles that form ordered intracellular clusters-as a model system to study the forces driving multiparticle assembly under cytoplasm-like conditions. Our calculations and experimental results show that the ordered assembly of GVs can be achieved by screening their mutual electrostatic repulsion with electrolytes and creating a crowding force with dissolved macromolecules. The precise balance of these forces results in different packing configurations. Biomacromolecules such as polylysine and DNA are capable of driving GV clustering. These results provide basic insights into how physically driven interactions affect the formation of protein superstructures, offer guidance for manipulating nanoparticle assembly in cellular environments through synthetic biology methods, and inform research on the biotechnology applications of GVs.
Topics: Cytoplasm; DNA; Macromolecular Substances; Nanoparticles; Static Electricity
PubMed: 34022233
DOI: 10.1016/j.bpj.2021.05.007 -
Foods (Basel, Switzerland) Jun 2021Tahini and tahini-based products are popular with consumers due to their special flavor and high nutritional values, but often have been linked to outbreaks. The...
Tahini and tahini-based products are popular with consumers due to their special flavor and high nutritional values, but often have been linked to outbreaks. The objective of this study was to compare effects of different kinds of natural antimicrobials on inactivation in undiluted and diluted tahini during thermal treatment and storage. Results showed that the Weibull model was more suitable to describe the thermal inactivation behavior of CICC21588 in two kinds of tahini than the first-order model. Inactivation curves were concave-upward in undiluted tahini but concave-downward in diluted tahini. During storage of undiluted tahini, 3% oregano oil caused extra 1.44 or 0.80 log CFU/g reductions after 7 days at 25 °C or 4 °C compared to the control and 0.5% citric acid caused an extra reduction of 0.75 log CFU/g after 7 d at 4 °C. For diluted tahini, 2-3% oregano oil and 0.4-0.5% ε-polylysine reduced more populations compared to undiluted tahini. These antimicrobials all inhibited the growth of during 24 h at 25 °C and ε-polylysine had the best effect. Furthermore, these antimicrobials enhanced the inactivation in diluted tahini during thermal treatment, and there was less of a synergistic effect of thermal and antimicrobials in undiluted tahini due to less sublethal injured cells caused by heat. This study may provide useful information for inactivation in tahini.
PubMed: 34204952
DOI: 10.3390/foods10061272 -
Bioactive Materials Apr 2024Wound management is an important issue that places enormous pressure on the physical and mental health of patients, especially in cases of infection, where the increased...
Wound management is an important issue that places enormous pressure on the physical and mental health of patients, especially in cases of infection, where the increased inflammatory response could lead to severe hypertrophic scars (HSs). In this study, a hydrogel dressing was developed by combining the high strength and toughness, swelling resistance, antibacterial and antioxidant capabilities. The hydrogel matrix was composed of a double network of polyvinyl alcohol (PVA) and agarose with excellent mechanical properties. Hyperbranched polylysine (HBPL), a highly effective antibacterial cationic polymer, and tannic acid (TA), a strong antioxidant molecule, were added to the hydrogel as functional components. Examination of antibacterial and antioxidant properties of the hydrogel confirmed the full play of the efficacy of HBPL and TA. In the studies of methicillin-resistant (MRSA) infection, the hydrogel had shown obvious promotion of wound healing, and more profoundly, significant suppression of scar formation. Due to the common raw materials and simple preparation methods, this hydrogel can be mass produced and used for accelerating wound healing while preventing HSs in infected wounds.
PubMed: 38261887
DOI: 10.1016/j.bioactmat.2023.12.019