Anionic Side Chains Research Articles

Investigation of the Nanoparticulation Method and Cell-Killing Effect following the Mitochondrial Delivery of Hydrophobic Porphyrin-Based Photosensitizers.

Photodynamic therapy is expected to be a less invasive treatment, and strategies for targeting mitochondria, the main sources of singlet oxygen, are attracting attention to increase the efficacy of photodynamic therapy and reduce its side effects. To date, we have succeeded in encapsulating the photosensitizer rTPA into MITO-Porter (MP), a mitochondria-targeted Drug Delivery System (DDS), aimed at mitochondrial delivery of the photosensitizer while maintaining its activity. In this study, we report the results of our studies to alleviate rTPA aggregation in an effort to improve drug efficacy and assess the usefulness of modifying the rTPA side chain to improve the mitochondrial retention of MITO-Porter, which exhibits high therapeutic efficacy. Conventional rTPA with anionic side chains and two rTPA analogs with side chains that were converted to neutral or cationic side chains were encapsulated into MITO-Porter. Low-MP (MITO-Porter with Low Drug/Lipid) exhibited high drug efficacy for all three types of rTPA, and in Low-MP, charged rTPA-encapsulated MP exhibited high drug efficacy. The cellular uptake and mitochondrial translocation capacities were similar for all particles, suggesting that differences in aggregation rates during the incorporation of rTPA into MITO-Porter resulted in differences in drug efficacy.

Mechanism of Protein Aggregation Inhibition by Arginine: Blockage of Anionic Side Chains Favors Unproductive Encounter Complexes.

Aggregation refers to the assembly of proteins into nonphysiological higher order structures. While amyloid has been studied extensively, much less is known about amorphous aggregation, a process that interferes with protein expression and storage. Free arginine (Arg+) is a widely used aggregation inhibitor, but its mechanism remains elusive. Focusing on myoglobin (Mb), we recently applied atomistic molecular dynamics (MD) simulations for gaining detailed insights into amorphous aggregation (Ng J. Phys. Chem. B 2021, 125, 13099). Building on that approach, the current work for the first time demonstrates that MD simulations can directly elucidate aggregation inhibition mechanisms. Comparative simulations with and without Arg+ reproduced the experimental finding that Arg+ significantly decreased the Mb aggregation propensity. Our data reveal that, without Arg+, protein-protein encounter complexes readily form salt bridges and hydrophobic contacts, culminating in firmly linked dimeric aggregation nuclei. Arg+ promotes the dissociation of encounter complexes. These "unproductive" encounter complexes are favored because Arg+ binding to D- and E- lowers the tendency of these anionic residues to form interprotein salt bridges. Side chain blockage is mediated largely by the guanidinium group of Arg+, which binds carboxylates through H-bond-reinforced ionic contacts. Our MD data revealed Arg+ self-association into a dynamic quasi-infinite network, but we found no evidence that this self-association is important for protein aggregation inhibition. Instead, aggregation inhibition by Arg+ is similar to that mediated by free guanidinium ions. The computational strategy used here should be suitable for the rational design of aggregation inhibitors with enhanced potency.

Enhancing the Features of DNA Mimic Foldamers for Structural Investigations.

DNA mimic foldamers based on aromatic oligoamide helices bearing anionic phosphonate side chains have been shown to bind to DNA-binding proteins sometimes orders of magnitude better than DNA itself. Here, we introduce new features in the DNA mimic foldamers to facilitate structural investigations of their interactions with proteins. Thirteen new foldamer sequences have been synthesized and characterized using NMR, circular dichroism, molecular modeling, and X-ray crystallography. The results show that foldamer helix handedness can be quantitatively biased by means of a single stereogenic center, that the foldamer structure can be made C2-symmetrical as in palindromic B-DNA sequences, and that associations between foldamer helices can be promoted utilizing dedicated C-terminal residues that act as sticky ends in B-DNA structures.

Biochemical and Biophysical Characterization of Tau and α-Linolenic Acid Vesicles In Vitro.

Alzheimer's disease (AD) is characterized by the abnormal accumulation of disordered protein, that is, extracellular senile plaques of amyloid-β (Aβ) and intracellular neurofibrillary tangles of Tau. Tau protein has gained the attention in recent years owing to the ability to propagate in a "prion-like" nature. The disordered protein Tau possesses a high positive charge, which allows its binding to anionic proteins and factors. The native disorder of proteins attends the β-sheet structure from its random-coiled conformation upon charge compensation by various polyanionic agents such as heparin, RNA, etc. Anionic lipids such as arachidonic acid (AA) and oleic acid (OA) are also one of the factors which can induce aggregation of Tau in physiological conditions. The free units of Tau protein can bind to lipid membranes through its repeat domain (RD), the anionic side chains of the membrane lipids induce aggregation of Tau by reducing the activation barrier. In this study, we investigated the role of α-linolenic acid (ALA) as an inducing agent for Tau aggregation in vitro conditions. Omega-3 fatty acids bear a capacity to reduce the pathology of Tau by downregulating the Tau phosphorylation pathway. We have studied by using various biochemical or biophysical methods the potency of ALA as an aggregating agent for Tau. We have implemented different techniques such as SDS-PAGE, transmission electron microscopy, CD spectroscopy to evaluated higher-order aggregates of Tau upon induction by ALA.

Plasmonic aptasensor with antifouling dual-functional surface layer for lysozyme detection in food

Antifouling coatings are critically necessary for optical biosensors for various analytical application sectors, from medical diagnostics to foodborne pathogen detection. They help avoid non-specific protein/cell attachment on the active biosensor surface and catch the analytes directly in the complex media. Advances in antifouling plasmonic surfaces have been mainly focused on detecting clinical biomarkers in real biofluids, whereas developing antifouling coatings for direct analysis of analytes in complex media has been scarcely investigated for food quality control and safety.Herein, we propose a new low-fouling poly-l-lysine (PLL)-based surface layer for directly detecting an allergen protein, lysozyme, in the food matrix using surface plasmon resonance. The PLL-based polymer contains densely immobilized anionic oligopeptide side chains to create an electric charge-balanced layer able to repel the non-specific adsorption of undesired molecules on the biosensor surface. It also includes sparsely attached aptamer probes for capturing lysozyme directly in food sources with no pre-analytical sample treatment. We optimized the surface layer fabrication condition and tested the dual-functional surface to evaluate its ability to detect the target protein selectively. The developed analytical approach allowed for achieving a limit of detection of 0.04 μg mL−1 (2.95 nM) and a limit of quantification of 0.13 μg mL−1 (8.95 nM). Lysozyme was successfully quantified in milk samples using the plasmonic dual-functional aptasensor without sample pre-treatment or target isolation, illustrating the device's utility.

Enhanced Organic Electrochemical Transistor Performance of Donor-Acceptor Conjugated Polymers Modified with Hybrid Glycol/Ionic Side Chains by Postpolymerization Modification.

Emergent bioelectronic technologies are underpinned by the organic electrochemical transistor (OECT), which employs an electrolyte medium to modulate the conductivity of its organic semiconductor channel. Here we utilize postpolymerization modification (PPM) on a conjugated polymer backbone to directly introduce glycolated or anionic side chains via fluoride displacement. The resulting polymers demonstrated increased volumetric capacitances, with subdued swelling, compared to their parent polymer in p-type enhancement mode OECTs. This increase in capacitance was attributed to their modified side chain configurations enabling cationic charge compensation for thin film electrochemical oxidation, as deduced from electrochemical quartz crystal microbalance measurements. An overall improvement in OECT performance was recorded for the hybrid glycol/ionic polymer compared to the parent, owing to its low swelling and bimodal crystalline orientation as imaged by grazing-incidence wide-angle X-ray scattering, enabling its high charge mobility at 1.02 cm2·V-1·s-1. Compromised device performance was recorded for the fully glycolated derivative compared to the parent, which was linked to its limited face-on stacking, which hindered OECT charge mobility at 0.26 cm2·V-1·s-1, despite its high capacitance. These results highlight the effectiveness of anionic side chain attachment by PPM as a means of increasing the volumetric capacitance of p-type conjugated polymers for OECTs, while retaining solid-state macromolecular properties that facilitate hole transport.

Anionic conductive group tunable amphoteric polybenzimidazole ion conductive membrane for vanadium redox flow battery

Anionic conductive group repels vanadium ions but is difficult to be incorporated into the selectively ionic conductive channels of the amphoteric ion conductive membrane with tunable ratio. Herein, the novel hydroxyl bromopropane anionic precursor side chain is proposed, in which the hydrophilic hydroxyl groups aggregate into ionic clusters with the sulfate ester cationic conductive side chains of polybenzimidazole via hydrogen bond during membrane casting. The subsequent conversion of the bromopropane precursor to quaternary ammonium can easily tune the anionic exchange capacity and integrate cations into the hydroxyl involved ionic conductive channels for selectively repelling vanadium ions. Compared with Nafion 212 membrane, the obtained amphoteric membrane exhibits much lower vanadium ion permeability (1.9 × 10−9 cm2s−1) and slightly lower area resistance (0.25 Ω cm2). Excellent performance of vanadium redox flow battery (coulombic efficiency, 99.2%, energy efficiency, 85.9% at 100 mA cm−2 and 0.31% discharge decay rate) is achieved, and efficiencies keep stable during the 500 cycles test. The performance of VRFB is surpass than that of most reported amphoteric ion conductive and Nafion 212 membranes.

Understanding the effect of an anionic side-chain on the nuclear spin dynamics of a polysaccharide

Understanding the effect of an anionic side-chain on the nuclear spin dynamics of a polysaccharide

Chryseobacterium cucumeris FHN1 keratinolytic enzyme valorized chicken feathers to amino acids with polar, anionic and non-polar imino side chain characteristics

Keratinolytic bacteria were isolated from poultry sludge samples using chicken feathers formulated media, and the identities of potent isolates were confirmed through 16S rRNA gene sequencing. Fermentation process conditions were optimized, and the amino acid profile of the feather hydrolysate was determined. Ten proteolytic bacteria were obtained from 20 isolates recovered from the samples with halo zone diameters on skimmed milk agar ranging from 15.5 ± 0.71 mm for PSW-15 to 28 ± 1.41 mm for PSW-08. The proteolytic bacteria showed percentage chicken feathers degradation that ranged from 29% for PSW-11 to 84% for PSW-14, and keratinase activity ranged from 99.99 ± 5.14 U/mL for PSW-15 to 761.82 ± 27.42 U/mL for PSW-14. The most potent isolates coded as PSW-14 and PSW-16 were identified as Chryseobacterium cucumeris FHN1 and Sphingobacterium multivorum HNFx, nucleotide sequences were submitted to the GenBank with accession numbers MW165487 and MW165488, respectively. C. cucumeris FHN1 showed the maximum keratinase activity of 1030.90 ± 15.42 U/mL after 72 h, at optimized fermentation conditions of initial medium pH 5–6, inoculum size (4%, v/v), chicken feather (1%, w/v), and incubation temperature (25 °C). The amino acid analysis showed serine (5.19%), aspartic acid (4.75%), glutamic acid (5.86%), and proline (6.08%) were relatively high in concentration and these amino acids possess polar, anionic and non-polar imino side chain characteristics. The chicken feathers-degrading capacity and the amino acid composition of the feather hydrolysate underpin C. cucumeris FHN1 dexterity with keratinous biomass valorization into valuable products with biotechnological application potentials.

Proton Switch in the Secondary Coordination Sphere to Control Catalytic Events at the Metal Center: Biomimetic Oxo Transfer Chemistry of Nickel Amidate Complex.

High-valent metal-oxo species are key intermediates for the oxygen atom transfer step in the catalytic cycles of many metalloenzymes. While the redox-active metal centers of such enzymes are typically supported by anionic amino acid side chains or porphyrin rings, peptide backbones might function as strong electron-donating ligands to stabilize high oxidation states. To test the feasibility of this idea in synthetic settings, we have prepared a nickel(II) complex of new amido multidentate ligand. The mononuclear nickel complex of this N5 ligand catalyzes epoxidation reactions of a wide range of olefins by using mCPBA as a terminal oxidant. Notably, a remarkably high catalytic efficiency and selectivity were observed for terminal olefin substrates. We found that protonation of the secondary coordination sphere serves as the entry point to the catalytic cycle, in which high-valent nickel species is subsequently formed to carry out oxo-transfer reactions. A conceptually parallel process might allow metalloenzymes to control the catalytic cycle in the primary coordination sphere by using proton switch in the secondary coordination sphere.

Unique and generic structural features of cholinium amino acid-based biocompatible ionic liquids.

Cholinium amino acid-based (Ch-AA) biocompatible ionic liquids (bio-ILs) are synthesized from renewable components and are efficiently used for biomass processing. However, their microscopic structural features that lead to their application as biomass solvents remain undetermined. Herein, we use atomistic simulations to investigate the structures of six different Ch-AA bio-ILs up to the nanometer length scale and demonstrate that, depending on the anion side chain structure, the respective IL exhibits structural ordering at different length scales. All the six Ch-AA bio-ILs investigated here show a generic feature of having a strong hydrogen bonding network between the hydroxyl group of the cholinium cation and the carboxyl group of the amino acid anions. We illustrate that each of these bio-ILs also displays a unique feature. Distinctive intermediate range structural ordering leads to heterogeneity in methioninate- and phenylalaninate-based ILs caused by the anion side chain segregation. Intermediate range ordering is not observed in glutaminate- and glutamate-based ILs because significant anion side chain and backbone interactions hinder the formation of side chain clusters. Interestingly, for the cysteinate-based IL, the side chains do not interact with the backbones and the intermediate range ordering is not observed because of a shorter anionic side chain.

Hybrid Sequences that Express both Aromatic Amide and α-Peptidic Folding Features.

Foldamers combining aliphatic and aromatic main‐chain units often produce atypical structures that cannot easily be accessed from purely aromatic or aliphatic sequences. We report solid‐state evidence that sequences comprising α‐amino acids and quinoline‐based monomers adopt conformations that combine the folding propensities of both components. Foldamers 2 and 3 having an XQQ repeat motif (X=α‐amino acid, Q=quinoline) were synthesized. Crystals of 2 (X=Phe, Q with an anionic side chain) obtained from water revealed an aromatic helix where amide groups belonging to the α‐amino acids created a hydrogen‐bond array typical of peptidic helices. Crystals of 3 (X=Ser, Q with a lipophilic side chain) obtained from organic solvents revealed a helix‐turn‐helix structure in which α‐amino acid side chains interfere with main‐chain hydrogen bonding. High sequence‐dependency of the conformation is typical of peptides but is shown here to include aromatic folding features.

Investigation into the molecular design and plasticizing effectiveness of HPEG-based polycarboxylate superplasticizers in alkali-activated slag

In this study, a series of HPEG PCE polymers with different anionic charge density and side chain lengths was synthesized via free-radical copolymerization from acrylic acid and α-methallyl poly (ethylene glycol) ether (HPEG) as macromonomer. The resulting PCE polymers were characterized via Size Exclusion Chromatography (SEC) and anionic charge titration. Thereafter, a correlation between their molecular structures and dispersing effectiveness was established for alkali-activated slag. A broad variety of HPEG PCE polymers was found to work in the alkali-activated slag (AAS) system. Finally, the working mechanism for best performing polymer was investigated via adsorption measurements. It was concluded that HPEG PCEs of high anionicity, high Mw and short side chain length adsorb particularity high amounts on slag, thus providing enhanced dispersion capacity as compared to other HPEG polymers.

Preparation of Poly(Ionic Liquid) Particles with Anionic Side Chain by Dispersion Polymerization.

Micrometer-sized poly(ionic liquid) (PIL) particles with an anionic side chain, poly(1-butyl-3-methylimidazolium 3-sulfopropyl methacrylate) (poly([Bmim][SPMT])), are successfully prepared by dispersion polymerization at 60°C in ethanol/ethyl acetate (2/8, w/w) with poly(vinylpyrrolidone) as a stabilizer. However, the obtained particles do not maintain the particulate state during drying at room temperature due to poly([Bmim][SPMT])'s low glass-transition temperature, below room temperature. In order to prevent coalescence and maintain particle shape, a cross-linking monomer is added after the nucleation stage of dispersion polymerization. Moreover, Fourier-transform infrared spectroscopy (FT-IR) demonstrates cation exchange between the obtained particles and metal cations (Ag+ ).

3D printable and injectable lactoferrin-loaded carboxymethyl cellulose-glycol chitosan hydrogels for tissue engineering applications.

In this study, carboxymethyl cellulose (CMC)-glycol chitosan (GC) hydrogel, a potential three-dimensional (3D) printing biomaterial ink for tissue engineering applications was synthesized using simple, biocompatible in situ-gelling Schiff's base reaction and ionic interactions. Different grades of hydrogels (C70G30, C50G50 and C30G70) were synthesized at physiological conditions. The oxidation of CMC and imine bond formation in the hydrogel were confirmed spectroscopically. Scanning electron microscopic images revealed the crosslinked interconnected pores in the cross-sectioned hydrogels (dried). Swelling (equilibrium: 1 h), porosity (~75%), in vitro degradation (>30 days) and thermal gravimetric analyses of the dried gels were studied. Initially, cytotoxicity assay was evaluated using mouse osteoblastic cells (MC3T3). These experiments revealed that CMC-GC gels formed stable hydrogel networks and were biocompatible. Particularly, C50G50 gels showed high printability (continuous extrusion) and post-printing stability (without secondary crosslinking). Gel 3D printing was optimized by varying the air pressure, temperature, needle size and nozzle speed, to obtain stable lattice structures (2 to 16 layers). The printed (2 and 5 layers) hydrogels showed high stability in phosphate buffer saline (PBS) solution (1 h), under UV light (1 h) and after autoclaving. The strut dimensions and porosity of the printed gels before and after the stability tests were analyzed. The hydrogel stability may be attributed to both the imine bond and ionic interaction between the cationic and anionic polymer side chains. Lactoferrin (glycoprotein) incorporated C50G50 gels showed sustained release up to 21 days in PBS (pH 7.4) solution and demonstrated increased biocompatibility (>80%) during in vitro cytotoxicity assays (MC3T3 cells and bone marrow mesenchymal stem cells) and Live/Dead assay (MC3T3 cells). A higher number of live osteoblast cells on the C50G50 hydrogels with increasing lactoferrin concentration was observed. These results show that the CMC-GC gels are promising bio-ink candidates for 3D printing and loading proteins or drugs for tissue engineering applications.

Structure and function of the calcium-selective TRP channel TRPV6.

Epithelial calcium channel TRPV6 is a member of the vanilloid subfamily of TRP channels that is permeable to cations and highly selective to Ca2+ ; it shows constitutive activity regulated negatively by Ca2+ and positively by phosphoinositol and cholesterol lipids. In this review, we describe the molecular structure of TRPV6 and discuss how its structural elements define its unique functional properties. High Ca2+ selectivity of TRPV6 originates from the narrow selectivity filter, where Ca2+ ions are directly coordinated by a ring of anionic aspartate side chains. Divalent cations Ca2+ and Ba2+ permeate TRPV6 pore according to the knock-off mechanism, while tight binding of Gd3+ to the aspartate ring blocks the channel and prevents Na+ from permeating the pore. The iris-like channel opening is accompanied by an α-to-π helical transition in the pore-lining transmembrane helix S6. As a result of this transition, the intracellular halves of the S6 helices bend and rotate by about 100deg, exposing different residues to the channel pore in the open and closed states. Channel opening is also associated with changes in occupancy of the transmembrane domain lipid binding sites. The inhibitor 2-aminoethoxydiphenyl borate (2-APB) binds to TRPV6 in a pocket formed by the cytoplasmic half of the S1-S4 transmembrane helical bundle and shifts open-closed channel equilibrium towards the closed state by outcompeting lipids critical for activation. Ca2+ inhibits TRPV6 via binding to calmodulin (CaM), which mediates Ca2+ -dependent inactivation. The TRPV6-CaM complex exhibits 1:1 stoichiometry; one TRPV6 tetramer binds both CaM lobes, which adopt a distinct head-to-tail arrangement. The CaM C-terminal lobe plugs the channel through a unique cation-π interaction by inserting the side chain of lysine K115 into a tetra-tryptophan cage at the ion channel pore intracellular entrance. Recent studies of TRPV6 structure and function described in this review advance our understanding of the role of this channel in physiology and pathophysiology and inform new therapeutic design.

Size-Selective Phagocytic Clearance of Fibrillar α-Synuclein through Conformational Activation of Complement Receptor 4.

Aggregation of α-synuclein (αSN) is an important histological feature of Parkinson disease. Recent studies showed that the release of misfolded αSN from human and rodent neurons is relevant to the progression and spread of αSN pathology. Little is known, however, about the mechanisms responsible for clearance of extracellular αSN. This study found that human complement receptor (CR) 4 selectively bound fibrillar αSN, but not monomeric species. αSN is an abundant protein in the CNS, which potentially could overwhelm clearance of cytotoxic αSN species. The selectivity of CR4 toward binding fibrillar αSN consequently adds an important αSN receptor function for maintenance of brain homeostasis. Based on the recently solved structures of αSN fibrils and the known ligand preference of CR4, we hypothesize that the parallel monomer stacking in fibrillar αSN creates a known danger-associated molecular pattern of stretches of anionic side chains strongly bound by CR4. Conformational change in the receptor regulated tightly clearance of fibrillar αSN by human monocytes. The induced change coupled concomitantly with phagolysosome formation. Data mining of the brain transcriptome in Parkinson disease patients supported CR4 as an active αSN clearance mechanism in this disease. Our results associate an important part of the innate immune system, namely complement receptors, with the central molecular mechanisms of CNS protein aggregation in neurodegenerative disorders.

Investigation on the optimal chemical structure of methacrylate ester based polycarboxylate superplasticizers to be used as cement grinding aid under laboratory conditions: Effect of anionicity, side chain length and dosage on grinding efficiency, mortar workability and strength development

Using polycarboxylate ether/ester (PCE) superplasticizers as cement grinding aid presents an interesting concept because the PCE could provide both grinding and fluidizing effect at the same time, thus achieving two benefits from the admixture. Here, a systematic study on the relationship between the molecular structure of ω-methoxy poly(ethylene glycol) methacrylate ester based PCE (MPEG-PCE) and their grinding efficiency was performed. A series of MPEG-PCEs with systematically modified compositions was employed and their impact on grinding efficiency was assessed. First of all, the PCE dosage required to achieve a certain fineness in a laboratory ball mill was determined. Then, the grinding efficiency of PCEs exhibiting different anionic charge density and side chain length was investigated. It was found that an optimal molar ratio of carboxylic acid : side chain exists, and that shorter side chains are more beneficial for grinding efficiency than longer ones. Furthermore, it was also found that intergrinding of MPEG-PCEs with clinker produces a cement with higher fluidity.

Impacts of the Terminal Charged Residues on Polyproline Conformation.

Cis-trans isomerization of proline is involved in various biological processes, such as protein folding, cell signaling, and ion-channel gating. Polyproline is a useful system for better understanding proline isomerization because it exists predominantly as two forms, all-cis polyproline I (PPI) and all-trans polyproline II (PPII) helices. The stability of PPI and PPII can be modulated by various effects, including aromatic-proline interactions, terminal charges, and stereoelectronic effects. Here, we used a series of oligoproline peptides in which positively charged or negatively charged amino acids were incorporated into the termini to investigate their effects on polyproline conformation. Circular dichroism measurements show that a cationic residue at the C-terminus or an anionic residue at the N-terminus increases the stability of a PPII helix; in particular, the C-terminal cationic residues impose an enormous impact on PPII stability. The electrostatic attractions between a cationic sidechain and the C-terminal carboxylate exhibit a greater effect than those between an anionic sidechain and the N-terminal ammonium on the conversion of PPII to PPI, suggesting that the stabilization effect of electrostatic interactions on PPII is directional. In contrast, incorporating a cationic residue seems more favorable than adding an anionic residue into the N-terminus because the cationic residue can stabilize PPI. Moreover, the predicted dipole moments from optimized oligopeptide models reveal that the macrodipole of the peptides with a cationic residue at the C-terminus exhibits the opposite direction to that of other peptides in the PPI conformation, suggesting that such a dipole distortion may cause these peptides to disfavor PPI helices. Together, we have found that the introduction of terminal electrostatic interactions can have a significant effect on PPII stability, providing useful information to the design of polyproline-based scaffolds for biomedical applications.

Effect of Ionic Functional Groups on the Oxidation State and Interfacial Binding Property of Catechol-Based Adhesive.

Adhesive hydrogels were prepared by copolymerizing dopamine methacrylamide with either acrylic acid (AAc) or N-(3-aminopropyl)methacrylamide hydrochloride (APMH). The effect of incorporating the anionic and cationic side chains on the oxidation state of catechol was characterized using the FOX assay to track the production of hydrogen peroxide byproduct generated during the autoxidation of catechol, and the interfacial binding property of the adhesive was determined by performing Johnson-Kendall-Roberts contact mechanics tests tested over a wide range of pH values (pH 3.0-9.0). The ionic species contributed to interfacial binding to surfaces with the opposite charge with measured work of adhesion values that were comparable to or in some cases higher than those of catechol. Addition of AAc minimized the oxidation of catechol even at a pH of 8.5 and correspondingly preserved the elevated adhesive property of catechol to both quartz and amine-functionalized surfaces. However, AAc lost its buffering capacity at pH 9.0, and catechol was oxidized at this pH. On the other hand, catechol formed a cohesive covalent bond with the network-bound amine side chain of APMH at basic pH, which interfered with the interfacial binding capability of APMH and the catechol.