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1.
Small ; 20(30): e2312041, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38438898

RESUMEN

Despite the considerable significance of utilizing ultra-thin film (utf) hydrogels as multipurpose platforms for biomedical applications, there is still an important lack of adequate characterization techniques suitable for such materials. In this Perspective, the use of quartz crystal microbalance with dissipation (QCM-D) coupled with spectral ellipsometry (SE) is presented as a potential tool for the complete characterization of utf-hydrogels due to its nanometric sensitivity and high versatility. Herein, the fundaments for utf-hydrogel characterization are settled down as far as the QCM-D/SE response is explored under a wide range of different in operando wet working conditions measurements such as temperature or liquid media, among others. Therefore, the design of measuring protocols capable to perform is proposed and compiled, for the first time, complete and precise characterization of the cross-link density, thickness variations (swelling ratio determination), stability analyses, and mechanical studies (including the simultaneous generation of stress-strain curves and the evaluation of the viscoelastic; leading to the final determination of the Poisson's ratio) under different in operando conditions. Finally, the future challenges and implications for the general characterization of soft-thin films are discussed.

2.
Protein Expr Purif ; 219: 106483, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38609025

RESUMEN

Mussel foot proteins (Mfps) possess unique binding properties to various surfaces due to the presence of L-3,4-dihydroxyphenylalanine (DOPA). Mytilus edulis foot protein-3 (Mefp-3) is one of several proteins in the byssal adhesive plaque. Its localization at the plaque-substrate interface approved that Mefp-3 plays a key role in adhesion. Therefore, the protein is suitable for the development of innovative bio-based binders. However, recombinant Mfp-3s are mainly purified from inclusion bodies under denaturing conditions. Here, we describe a robust and reproducible protocol for obtaining soluble and tag-free Mefp-3 using the SUMO-fusion technology. Additionally, a microbial tyrosinase from Verrucomicrobium spinosum was used for the in vitro hydroxylation of peptide-bound tyrosines in Mefp-3 for the first time. The highly hydroxylated Mefp-3, confirmed by MALDI-TOF-MS, exhibited excellent adhesive properties comparable to a commercial glue. These results demonstrate a concerted and simplified high yield production process for recombinant soluble and tag-free Mfp3-based proteins with on demand DOPA modification.


Asunto(s)
Dihidroxifenilalanina , Mytilus edulis , Animales , Dihidroxifenilalanina/química , Dihidroxifenilalanina/metabolismo , Mytilus edulis/genética , Mytilus edulis/química , Mytilus edulis/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Verrucomicrobia/genética , Verrucomicrobia/metabolismo , Monofenol Monooxigenasa/genética , Monofenol Monooxigenasa/metabolismo , Monofenol Monooxigenasa/química , Proteínas/genética , Proteínas/química , Proteínas/aislamiento & purificación , Hidroxilación , Escherichia coli/genética , Escherichia coli/metabolismo
3.
Environ Sci Technol ; 2024 Jan 22.
Artículo en Inglés | MEDLINE | ID: mdl-38247403

RESUMEN

The mobility and distribution of heavy metal ions (HMs) in aquatic environments are significantly influenced by humic acid (HA), which is ubiquitous. A quantitative understanding of the interaction mechanism underlying the adsorption and retention of HMs by HA is of vital significance but remains elusive. Herein, the interaction mechanism between HA and different types of HMs (i.e., Cd(II), Pb(II), arsenate, and chromate) was quantitatively investigated at the nanoscale. Based on quartz crystal microbalance with dissipation tests, the adsorption capacities of Pb(II), Cd(II), As(V), and Cr(VI) ionic species on the HA surface were measured as ∼0.40, ∼0.25, ∼0.12, and ∼0.02 nmol cm-2, respectively. Atomic force microscopy force results showed that the presence of Pb(II)/Cd(II) cations suppressed the electrostatic double-layer repulsion during the approach of two HA surfaces and the adhesion energy during separation was considerably enhanced from ∼2.18 to ∼5.05/∼4.18 mJ m-2. Such strong adhesion stems from the synergistic metal-HA complexation and cation-π interaction, as evidenced by spectroscopic analysis and theoretical simulation. In contrast, As(V)/Cr(VI) oxo-anions could form only weak hydrogen bonds with HA, resulting in similar adhesion energies for HA-HA (∼2.18 mJ m-2) and HA-As(V)/Cr(VI)-HA systems (∼2.26/∼1.96 mJ m-2). This work provides nanoscale insights into quantitative HM-HA interactions, improving the understanding of HMs biogeochemical cycling.

4.
J Sci Food Agric ; 2024 Feb 03.
Artículo en Inglés | MEDLINE | ID: mdl-38308593

RESUMEN

BACKGROUND: Polyphenols are a group of compounds found in grapes, musts, and wines. Their levels are crucial for grape ripening, proper must fermentation, and final wine characteristics. Standard chemical analysis is commonly used to detect these compounds, but it is costly, time consuming, and requires specialized laboratories and operators. To address this, this study explores a functionalized acoustic sensor for detecting oenological polyphenols. RESULTS: The method involves utilizing a quartz crystal microbalance with dissipation monitoring (QCM-D) to detect the target analyte by using a gelatin-based probe layer. The sensor is functionalized by optimizing the probe coverage density to maximize its performance. This is achieved by using 12-mercaptododecanoic acid (12-MCA) to immobilize the probe onto the gold sensor surface, and dithiothreitol (DTT) as a reducing and competitive binding agent. The concentration of 12-MCA and DTT in the solutions is varied to control the probe density. QCM-D measurements demonstrate that the probe density can be effectively adjusted using this approach, ranging from 0.2 × 1013 to 2 × 1013 molecules cm-2 . This study also investigates the interaction between the probe and tannins, confirming the ability of the sensor to detect them. Interestingly, the lower probe coverage achieves higher detection signals when normalized to probe immobilization signals. Moreover, significant changes in mechanical properties of the functionalization layer are observed after the interaction with samples. CONCLUSION: The combination of QCM-D with gelatin functionalization holds great promise for future applications in the wine industry. It offers real-time monitoring capabilities, requires minimal sample preparation, and provides high sensitivity for quality control purposes. © 2024 Society of Chemical Industry.

5.
Small ; 19(23): e2207125, 2023 06.
Artículo en Inglés | MEDLINE | ID: mdl-36899445

RESUMEN

Membrane fusion is essential for the basal functionality of eukaryotic cells. In physiological conditions, fusion events are regulated by a wide range of specialized proteins, operating with finely tuned local lipid composition and ionic environment. Fusogenic proteins, assisted by membrane cholesterol and calcium ions, provide the mechanical energy necessary to achieve vesicle fusion in neuromediator release. Similar cooperative effects must be explored when considering synthetic approaches for controlled membrane fusion. We show that liposomes decorated with amphiphilic Au nanoparticles (AuLips) can act as minimal tunable fusion machinery. AuLips fusion is triggered by divalent ions, while the number of fusion events dramatically changes with, and can be finely tuned by, the liposome cholesterol content. We combine quartz-crystal-microbalance with dissipation monitoring (QCM-D), fluorescence assays, and small-angle X-ray scattering (SAXS) with molecular dynamics (MD) at coarse-grained (CG) resolution, revealing new mechanistic details on the fusogenic activity of amphiphilic Au nanoparticles (AuNPs) and demonstrating the ability of these synthetic nanomaterials to induce fusion regardless of the divalent ion used (Ca2+ or Mg2+ ). The results provide a novel contribution to developing new artificial fusogenic agents for next-generation biomedical applications that require tight control of the rate of fusion events (e.g., targeted drug delivery).


Asunto(s)
Liposomas , Nanopartículas del Metal , Oro , Dispersión del Ángulo Pequeño , Difracción de Rayos X , Proteínas , Colesterol , Iones
6.
Environ Sci Technol ; 57(39): 14707-14716, 2023 10 03.
Artículo en Inglés | MEDLINE | ID: mdl-37722069

RESUMEN

Plastic fate in freshwater systems is dependent on particle size, morphology, and physicochemical surface properties (e.g., charge, surface roughness, and hydrophobicity). Environmental aging processes, such as photochemical weathering and eco-corona formation due to dissolved organic matter (DOM) adsorption on plastic surfaces, can alter their physicochemical properties, affecting fate and transport. While plastic aging has been studied from a materials science perspective, its specific implications in environmental contexts remain less understood. Although photochemical weathering and eco-corona formation occur simultaneously in the environment, in this work, we systematically assessed the effects of photochemical weathering on the physicochemical properties of polymers (polyethylene, polypropylene, polyethylene terephthalate, and polystyrene) and how this influences the adsorption of DOMs (Suwannee River humic acid, fulvic acid, and natural organic matter) relative to pristine polymers. Pristine polymers initially had different and distinct physicochemical surface properties, but upon aging, they became more similar in terms of surface properties. Photochemical weathering resulted in a decrease in polymer film thickness, an increase in surface roughness, and hydrophilicity. DOM adlayers on the polymer surfaces resulted in more comparable wettability, effectively masking the initial polymer properties. Collectively, this study explores the physiochemical changes polymers undergo in laboratory studies mimicking environmental conditions. Understanding these changes is the initial step to rationalizing and predicting processes and interactions such as heteroaggregation that dictate the fate of plastics in the environment.


Asunto(s)
Materia Orgánica Disuelta , Polímeros , Polímeros/química , Adsorción , Poliestirenos , Sustancias Húmicas/análisis , Plásticos
7.
Sensors (Basel) ; 23(3)2023 Jan 25.
Artículo en Inglés | MEDLINE | ID: mdl-36772387

RESUMEN

Quartz crystal microbalance with dissipation monitoring (QCM-D) is a well-established technique for studying soft films. It can provide gravimetric as well as nongravimetric information about a film, such as its thickness and mechanical properties. The interpretation of sets of overtone-normalized frequency shifts, ∆f/n, and overtone-normalized shifts in half-bandwidth, ΔΓ/n, provided by QCM-D relies on a model that, in general, contains five independent parameters that are needed to describe film thickness and frequency-dependent viscoelastic properties. Here, we examine how noise inherent in experimental data affects the determination of these parameters. There are certain conditions where noise prevents the reliable determination of film thickness and the loss tangent. On the other hand, we show that there are conditions where it is possible to determine all five parameters. We relate these conditions to the mathematical properties of the model in terms of simple conceptual diagrams that can help users understand the model's behavior. Finally, we present new open source software for QCM-D data analysis written in Python, PyQTM.

8.
Int J Mol Sci ; 24(17)2023 Aug 25.
Artículo en Inglés | MEDLINE | ID: mdl-37686023

RESUMEN

The profound understanding and detailed evaluation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (SCoV2-S) protein and specific antibody interaction mechanism is of high importance in the development of immunosensors for COVID-19. In the present work, we studied a model system of immobilized SCoV2-S protein and specific monoclonal antibodies by molecular dynamics of immune complex formation in real time. We simultaneously applied spectroscopic ellipsometry and quartz crystal microbalance with dissipation to reveal the features and steps of the immune complex formation. We showed direct experimental evidence based on acoustic and optical measurements that the immune complex between covalently immobilized SCoV2-S and specific monoclonal antibodies is formed in two stages. Based on these findings it was demonstrated that applying a two-step binding mathematical model for kinetics analysis leads to a more precise determination of interaction rate constants than that determined by the 1:1 Langmuir binding model. Our investigation showed that the equilibrium dissociation constants (KD) determined by a two-step binding model and the 1:1 Langmuir model could differ significantly. The reported findings can facilitate a deeper understanding of antigen-antibody immune complex formation steps and can open a new way for the evaluation of antibody affinity towards corresponding antigens.


Asunto(s)
Técnicas Biosensibles , COVID-19 , Humanos , Glicoproteína de la Espiga del Coronavirus , Complejo Antígeno-Anticuerpo , Afinidad de Anticuerpos , Inmunoensayo , SARS-CoV-2 , Anticuerpos Monoclonales
9.
Molecules ; 28(11)2023 May 29.
Artículo en Inglés | MEDLINE | ID: mdl-37298891

RESUMEN

This study investigates the interaction between montmorillonite and polyacrylamide (PAM) with different ionic types using quartz crystal microbalance with dissipation monitoring (QCM-D) and molecular dynamics (MD) simulations. The goal was to understand the effect of ionicity and ionic type on polymer deposition on montmorillonite surfaces. The results of the QCM-D analysis showed that a decrease in pH led to an increase in the adsorption of montmorillonite on the alumina surface. The ranking of adsorption mass on alumina and pre-adsorbed montmorillonite alumina surfaces was found to be cationic polyacrylamide (CPAM) > polyacrylamide (NPAM) > anionic polyacrylamide (APAM). The study also found that CPAM had the strongest bridging effect on montmorillonite nanoparticles, followed by NPAM, while APAM had a negligible bridging effect. The MD simulations showed that ionicity had a significant influence on the adsorption of polyacrylamides. The cationic functional group N(CH3)3+ had the strongest attraction interaction with the montmorillonite surface, followed by the hydrogen bonding interaction of the amide functional group CONH2, and the anionic functional group COO- had a repulsive interaction. The results suggest that at high ionicity levels, CPAM can be adsorbed on the montmorillonite surface, while at low ionicity levels, APAM may still be adsorbed with a strong coordination trend.


Asunto(s)
Bentonita , Tecnicas de Microbalanza del Cristal de Cuarzo , Adsorción , Tecnicas de Microbalanza del Cristal de Cuarzo/métodos , Simulación de Dinámica Molecular , Iones , Óxido de Aluminio , Propiedades de Superficie
10.
Molecules ; 28(14)2023 Jul 22.
Artículo en Inglés | MEDLINE | ID: mdl-37513453

RESUMEN

The development of novel drug delivery systems (DDSs) with promising antibacterial properties is essential for facing the emergency of increasing resistance to antimicrobial agents. The antibacterial features of quercetin and its metal complexes have been broadly investigated. However, several drawbacks affect their activity and effectiveness. In this work, we propose a DDS based on a pH-responsive cobalt(II)-coordinated assembly containing quercetin and polyacrylic acid. This system is suggested to trigger the release of the model drug in a pH-dependent mode by exploiting the localized acidic environment at the bacterial infection sites under anaerobic conditions. The delivery system has been designed by accurately examining the species and the multiple equilibria occurring in solution among the assembly components. The formation of cobalt(II) complexes with quercetin in the absence or presence of the pH-responsive polyacrylic acid was investigated in buffered aqueous solution at pH 7.4 using spectrophotometric (UV-Vis) and calorimetric (ITC) techniques. The determined binding affinities and thermodynamic parameters that resulted are essential for the development of a DDS with improved binding and release capabilities. Furthermore, the affinity of the polymer-cobalt(II) complex toward the model antimicrobial flavonoid was explored at the solid-liquid interface by quartz crystal microbalance (QCM-D) experiments, which provided marked evidence for drug loading and release under pH control.


Asunto(s)
Complejos de Coordinación , Quercetina , Quercetina/farmacología , Cobalto/química , Complejos de Coordinación/farmacología , Complejos de Coordinación/química , Antibacterianos/farmacología , Antibacterianos/química , Concentración de Iones de Hidrógeno
11.
Chemphyschem ; 23(3): e202100790, 2022 02 04.
Artículo en Inglés | MEDLINE | ID: mdl-34850511

RESUMEN

There is still a lack of deep understanding on the reaction kinetics and mechanism of thiol etching of gold. Herein, by using the sensor of quartz crystal microbalance (QCM) as the sacrificial probe, the etching reaction of gold has been studied by employing cysteamine (CS) as a typical thiol etchant. The etching reaction is verified as diffusion-controlled and shows a half-order reaction kinetics. It is demonstrated that intact thiol and amino on CS are both crucial for its etching ability to gold. Applied potentials can affect the electron transfer and hence can be used to regulate the gold etching. Our results also reveal that only two carbon atoms of the spacer between thiol and amino on CS are very critical to the excellent etching ability. This work exhibits a new route to explore the thiol etching reaction of gold and elucidates the reaction kinetics and mechanism.


Asunto(s)
Técnicas Biosensibles , Tecnicas de Microbalanza del Cristal de Cuarzo , Técnicas Biosensibles/métodos , Oro/química , Tecnicas de Microbalanza del Cristal de Cuarzo/métodos , Compuestos de Sulfhidrilo/química
12.
Environ Sci Technol ; 56(20): 14763-14773, 2022 10 18.
Artículo en Inglés | MEDLINE | ID: mdl-36197031

RESUMEN

Extracellular polymeric substances (EPSs) can conform and orient on the surface according to the applied aquatic conditions. While pH elevation usually removes EPSs from membranes, small changes in pH can change the adsorbed EPS conformation and orientation, resulting in a decrease in membrane permeability. Accordingly, EPS layers were tested with localized surface plasmon resonance (LSPR) sensing and quartz crystal microbalance with dissipation monitoring (QCM-D) using a hybrid sensor. A novel membrane-mimetic hybrid QCM-D-LSPR sensor was designed to indicate both "dry" mass and mechanical load ("wet" mass) of the adsorbed EPS. The effect of pH on the EPS layer's viscoelastic properties and hydrated thickness analyzed by QCM-D corroborates with the shift in EPS areal concentration, ΓS, and the associated EPS conformation, analyzed by LSPR. As pH elevates, the processes of (i) elevation in EPS layer's thickness (QCM-D) and (ii) decrease in the EPS areal density, ΓS (LSPR), provide a clear indication for changes in EPS conformation, which decrease the effective ultrafiltration (UF) membrane pore diameter. This decrease in the pore diameter together with the increase in surface hydrophobicity elevates UF membrane hydraulic resistance.


Asunto(s)
Matriz Extracelular de Sustancias Poliméricas , Ultrafiltración , Adsorción , Concentración de Iones de Hidrógeno , Resonancia por Plasmón de Superficie
13.
Sensors (Basel) ; 22(15)2022 Jul 31.
Artículo en Inglés | MEDLINE | ID: mdl-35957285

RESUMEN

In this paper, an FPGA (Field Programmable Gate Array)-based digital architecture for the measurement of quartz crystal microbalance (QCM) oscillating frequency of transient responses, i.e., in QCM-D (QCM and Dissipation) applications, is presented. The measurement system is conceived for operations in liquid, with short QCM transient responses due to the large mechanical load. The proposed solution allows for avoiding the complex processing systems typically required by the QCM-D techniques and grants frequency resolutions better than 1 ppm. The core of the architecture is a reciprocal digital frequency meter, combined with the preprocessing of the QCM signal through mixing operations, such as a step-down of the input frequency and reducing the measurement error. The measurement error is further reduced through averaging. Different strategies are proposed to implement the proposed measurement solution, comprising an all-digital circuit and mixed analog/digital ones. The performance of the proposed architectures is theoretically derived, compared, and analyzed by means of experimental data obtained considering 10 MHz QCMs and 200 µs long transient responses. A frequency resolution of about 240 ppb, which corresponds to a Sauerbrey mass resolution of 8 ng/cm2, is obtained for the all-digital solution, whereas for the mixed solution the resolution halves to 120 ppb, with a measurement time of about one second over 100 repetitions.


Asunto(s)
Técnicas Biosensibles , Tecnicas de Microbalanza del Cristal de Cuarzo , Técnicas Biosensibles/métodos , Cuarzo/química
14.
Environ Res ; 194: 110646, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-33359458

RESUMEN

Anammox bacteria are widely found to grow in bioaggregates form, but the reason for their high aggregation ability remains elusive. In this study, four kinds of sludge, i.e., anammox granules, anaerobic granules, aerobic granules, and partial nitrification flocs, were studied and compared to investigate their differences in adherence properties. We directly explored the adherence properties of sludge samples before and after extracellular polymeric substances (EPS) extraction, using quartz-crystal microbalance technique with dissipation monitoring technique. Results showed that EPS indeed stimulated the adherence properties of all sludge samples. The most striking feature here is that anammox consortia had the highest adhesion rate and mass, and formed the most compact layer on the gold-coated sensor surfaces both before and after EPS extraction among the four sludge samples, indicating their inherent high adhesion ability. The composition and spectral characteristics of EPS samples were also investigated, and it reveals that the relatively high extracellular proteins/polysaccharides ratio of anammox granules (3.2 ± 0.4) rather than total EPS concentration had contributed to their high adhesion ability. The findings are helpful for understanding the adherence properties of anammox bacteria, and will serve as a guide for further researches to exploring the aggregation process of anammox bacteria.


Asunto(s)
Reactores Biológicos , Nitrógeno , Matriz Extracelular de Sustancias Poliméricas , Nitrificación , Aguas del Alcantarillado
15.
Lett Appl Microbiol ; 73(2): 257-267, 2021 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-34028067

RESUMEN

Legionella pneumophila cell surface hydrophobicity and charge are important determinants of their mobility and persistence in engineered water systems (EWS). These surface properties may differ depending on the growth phase of L. pneumophila resulting in variable adhesion and persistence within EWS. We describe the growth-dependent variations in L. pneumophila cell surface hydrophobicity and surface charge using the microbial adhesion to hydrocarbon assay and microelectrophoresis, respectively, and their role in cell adhesion to stainless steel using a quartz crystal microbalance with dissipation (QCM-D) monitoring instrument. We observed a steady increase in L. pneumophila hydrophobicity during their lifecycle in culture media. Cell surfaces of stationary phase L. pneumophila were significantly more hydrophobic than their lag and midexponential counterparts. No significant changes in L. pneumophila cell surface charge were noted. Morphology of L. pneumophila remained relatively constant throughout their lifecycle. In the QCM-D study, lag and exponential phase L. pneumophila weakly adhered to stainless steel surfaces resulting in viscoelastic layers. In contrast, stationary phase bacteria were tightly and irreversibly bound to the surfaces, forming rigid layers. Our results suggest that the stationary phase of L. pneumophila would highly favour their adhesion to plumbing surfaces and persistence in EWS.


Asunto(s)
Adhesión Bacteriana , Legionella pneumophila/fisiología , Tecnicas de Microbalanza del Cristal de Cuarzo , Acero Inoxidable , Interacciones Hidrofóbicas e Hidrofílicas , Legionella pneumophila/ultraestructura , Microscopía Electrónica de Rastreo , Propiedades de Superficie
16.
Food Hydrocoll ; 111: 106364, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33536697

RESUMEN

The aim of this work was to compare the surface adsorption and lubrication properties of plant and dairy proteins. Whey protein isolate (WPI) and pea protein isolate (PPI) were chosen as model animal and plant proteins, respectively, and various protein concentrations (0.1-100 mg/mL) were studied with/without heat treatment (90 °C/60 min). Quartz crystal microbalance with dissipation monitoring (QCM-D) experiments were performed on hydrophilic (gold) and hydrophobic polydimethylsiloxane (PDMS) sensors, with or without a mucin coating, latter was used to mimic the oral surface. Soft tribology using PDMS tribopairs in addition to wettability measurements, physicochemical characterization (size, charge, solubility) and gel electrophoresis were performed. Soluble fractions of PPI adsorbed to significantly larger extent on PDMS surfaces, forming more viscous films as compared to WPI regardless of heat treatment. Introducing a mucin coating on a PDMS surface led to a decrease in binding of the subsequent dietary protein layers, with PPI still adsorbing to a larger extent than WPI. Such large hydrated mass of PPI resulted in superior lubrication performance at lower protein concentration (≤10 mg/mL) as compared to WPI. However, at 100 mg/mL, WPI was a better lubricant than PPI, with the former showing the onset of elastohydrodynamic lubrication. Enhanced lubricity upon heat treatment was attributed to the increase in apparent viscosity. Fundamental insights from this study reveal that pea protein at higher concentrations demonstrates inferior lubricity than whey protein and could result in unpleasant mouthfeel, and thus may inform future replacement strategies when designing sustainable food products.

17.
Sensors (Basel) ; 21(10)2021 May 17.
Artículo en Inglés | MEDLINE | ID: mdl-34067761

RESUMEN

The response of the quartz crystal microbalance (QCM, also: QCM-D for "QCM with Dissipation monitoring") to loading with a diverse set of samples is reviewed in a consistent frame. After a brief introduction to the advanced QCMs, the governing equation (the small-load approximation) is derived. Planar films and adsorbates are modeled based on the acoustic multilayer formalism. In liquid environments, viscoelastic spectroscopy and high-frequency rheology are possible, even on layers with a thickness in the monolayer range. For particulate samples, the contact stiffness can be derived. Because the stress at the contact is large, the force is not always proportional to the displacement. Nonlinear effects are observed, leading to a dependence of the resonance frequency and the resonance bandwidth on the amplitude of oscillation. Partial slip, in particular, can be studied in detail. Advanced topics include structured samples and the extension of the small-load approximation to its tensorial version.

18.
Int J Mol Sci ; 22(3)2021 Jan 28.
Artículo en Inglés | MEDLINE | ID: mdl-33525751

RESUMEN

The physicochemical properties of protein layers at the solid-liquid interface are essential in many biological processes. This study aimed to link the structural analysis of adsorbed lysozyme at the water/gold surface at pH 7.5 in a wide range of concentrations. Particular attention was paid to the protein's structural stability and the hydration of the protein layers formed at the interface. Complementary methods such as multi-parameter surface plasmon resonance (MP-SPR), quartz crystal microbalance with energy dissipation (QCM-D), and infrared spectroscopy (FTIR) were used for this purpose. The MP-SPR and QCM-D studies showed that, during the formation of a monolayer on the gold surface, the molecules' orientation changes from side-on to end-on. In addition, bilayer formation is observed when adsorbing in the high-volume concentration range >500 ppm. The degree of hydration of the monolayer and bilayer varies depending on the degree of surface coverage. The hydration of the system decreases with filling the layer in both the monolayer and the bilayer. Hydration for the monolayer varies in the range of 50-70%, because the bilayer is much higher than 80%. The degree of hydration of the adsorption layer has a crucial influence on the protein layers' viscoelastic properties. In general, an increase in the filling of a layer is characterized by a rise in its rigidity. The use of infrared spectroscopy allowed us to determine the changes taking place in the secondary structure of lysozyme due to its interaction with the gold surface. Upon adsorption, the content of II-structures corresponding to ß-turn and random lysozyme structures increases, with a simultaneous decrease in the content of the ß-sheet. The increase in the range of ß-turn in the structure determines the lysozyme structure's stability and prevents its aggregation.


Asunto(s)
Oro/química , Muramidasa/química , Agua/química , Adsorción , Concentración de Iones de Hidrógeno , Estructura Secundaria de Proteína , Tecnicas de Microbalanza del Cristal de Cuarzo , Espectroscopía Infrarroja por Transformada de Fourier , Resonancia por Plasmón de Superficie , Propiedades de Superficie
19.
Appl Environ Microbiol ; 86(23)2020 11 10.
Artículo en Inglés | MEDLINE | ID: mdl-32978123

RESUMEN

The outer membrane c-type cytochromes (c-Cyts) OmcA and MtrC in Shewanella are key terminal reductases that bind and transfer electrons directly to iron (hydr)oxides. Although the amounts of OmcA and MtrC at the cell surface and their molecular structures are largely comparable, MtrC is known to play a more important role in dissimilatory iron reduction. To explore the roles of these outer membrane c-Cyts in the interaction of Shewanella oneidensis MR-1 with iron oxides, the processes of attachment of S. oneidensis MR-1 wild type and c-type cytochrome-deficient mutants (the ΔomcA, ΔmtrC, and ΔomcA ΔmtrC mutants) to goethite are compared via quartz crystal microbalance with dissipation monitoring (QCM-D). Strains with OmcA exhibit a rapid initial attachment. The quantitative model for QCM-D responses reveals that MtrC enhances the contact area and contact elasticity of cells with goethite by more than one and two times, respectively. In situ attenuated total reflectance Fourier transform infrared two-dimensional correlation spectroscopic (ATR-FTIR 2D-CoS) analysis shows that MtrC promotes the initial interfacial reaction via an inner-sphere coordination. Atomic force microscopy (AFM) analysis demonstrates that OmcA enhances the attractive force between cells and goethite by about 60%. As a result, OmcA contributes to a higher attractive force with goethite and induces a rapid short-term attachment, while MtrC is more important in the longer-term interaction through an enhanced contact area, which promotes interfacial reactions. These results reveal that c-Cyts OmcA and MtrC adopt different mechanisms for enhancing the attachment of S. oneidensis MR-1 cells to goethite. It improves our understanding of the function of outer membrane c-Cyts and the influence of cell surface macromolecules in cell-mineral interactions.IMPORTANCEShewanella species are one group of versatile and widespread dissimilatory iron-reducing bacteria, which are capable of respiring insoluble iron minerals via six multiheme c-type cytochromes. Outer membrane c-type cytochromes (c-Cyts) OmcA and MtrC are the terminal reductases in this pathway and have comparable protein structures. In this study, we elucidate the different roles of OmcA and MtrC in the interaction of S. oneidensis MR-1 with goethite at the whole-cell level. OmcA confers enhanced affinity toward goethite and results in rapid attachment. Meanwhile, MtrC significantly increases the contact area of bacterial cells with goethite and promotes the interfacial reaction, which may explain its central role in extracellular electron transfer. This study provides novel insights into the role of bacterial surface macromolecules in the interfacial interaction of bacteria with minerals, which is critical to the development of a comprehensive understanding of cell-mineral interactions.


Asunto(s)
Adhesión Bacteriana/genética , Proteínas Bacterianas/genética , Grupo Citocromo c/genética , Compuestos de Hierro/metabolismo , Minerales/metabolismo , Shewanella/genética , Proteínas Bacterianas/metabolismo , Grupo Citocromo c/metabolismo , Shewanella/metabolismo
20.
Curr Top Membr ; 86: 1-55, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-33837691

RESUMEN

Lipid membranes are involved in many physiological processes like recognition, signaling, fusion or remodeling of the cell membrane or some of its internal compartments. Within the cell, they are the ultimate barrier, while maintaining the fluidity or flexibility required for a myriad of processes, including membrane protein assembly. The physical properties of in vitro model membranes as model cell membranes have been extensively studied with a variety of techniques, from classical thermodynamics to advanced modern microscopies. Here we review the nanomechanics of solid-supported lipid membranes with a focus in their phase behavior. Relevant information obtained by quartz crystal microbalance with dissipation monitoring (QCM-D) and atomic force microscopy (AFM) as complementary techniques in the nano/mesoscale interface is presented. Membrane morphological and mechanical characterization will be discussed in the framework of its phase behavior, phase transitions and coexistence, in simple and complex models, and upon the presence of cholesterol.


Asunto(s)
Membrana Dobles de Lípidos , Tecnicas de Microbalanza del Cristal de Cuarzo , Membrana Celular , Colesterol , Microscopía de Fuerza Atómica
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