Acid Conformation Research Articles

Intermolecular Hydrogen-Bonded Interactions of Oxalic Acid Conformers with Sulfuric Acid and Ammonia.

Oxalic acid is one of the simplest naturally occurring dicarboxylic acids that is abundantly found in the atmosphere, and it has several stable structural conformers. Hydrogen-bonded interactions of oxalic acid with other atmospheric molecules are important, as they might influence the chemical composition of the atmosphere, thereby impacting atmospheric chemistry and environmental processes. In this work, we used density functional calculations with the M06-2X/6-311++G(3df,3pd) model to examine the interaction of five oxalic acid conformers with sulfuric acid and ammonia-two widely recognized atmospheric nucleation precursor molecules-with the aim of observing the hydrogen-bonding characteristics of the conformers individually. An extensive and systematic quantum-chemical calculation has been conducted to analyze the structural, thermodynamical, electrical, and spectroscopic characteristics of several binary and ternary clusters mediated by five oxalic acid conformers. Our analysis of the electronic-binding energies and free energy changes associated with the formation of the clusters at ambient temperature reveals that multiple conformations of oxalic acid have the potential to engage in stable cluster formation in the atmosphere. In fact, the highest energy oxalic acid conformer exhibits the lowest bonding free energy in most cases. According to our calculations, clusters of oxalic acid with sulfuric acid demonstrate greater thermodynamic stability, a higher probability of formation, and more intense light scattering compared to clusters with ammonia. Furthermore, the analysis of successive cluster formation reveals that clusters formed between sulfuric and oxalic acids are more likely to grow spontaneously than those formed between ammonia and oxalic acid.

PH modulates efficiency of singlet oxygen production by flavin cofactors.

Flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN) are frequently used interchangeably in the catalysis of various reactions as part of flavoenzymes because they have the same functional component, the isoalloxazine ring. However, they differ significantly in their conformational properties. The inclusion of two planar rings in the structure of FAD greatly increases the range of possible conformations compared to FMN. An exemplary instance of this is the remarkable disparity in singlet oxygen efficiency production, Φ Δ, between FMN and FAD. Under neutral pH conditions, FAD has low photosensitizing activity with Φ Δ ∼ 0.07 while FMN demonstrates high photosensitizing activity with Φ Δ ∼ 0.6. Both adenine rings and isoalloxazine in FAD contain pH titratable groups. Through comprehensive analysis of the kinetics of the transient absorbance of the triplet state and the phosphorescence of singlet oxygen from FAD and FMN, we determined the correlation between different conformational states and the pH-dependent generation of singlet oxygen. Based on our findings, we may deduce that within the pH range of pH 2 to pH 13, only two out of the five potential structural states of FAD are capable of efficiently producing singlet oxygen. There are two open conformations: (i) an acidic FAD conformation with a protonated adenine ring, which is around 10 times more populated than the neutral open FAD conformation, and (ii) a neutral pH FAD conformation, which is significantly less populated. The FAD conformer with a protonated adenine ring at acidic pH generates singlet oxygen with approximately 50% efficiency compared to the constantly open FMN at neutral pH. This may have implications for singlet oxygen synthesis in acidic environments.

Solution NMR backbone assignment of the N-terminal tandem Zα1-Zα2 domains of Z-DNA binding protein 1.

The detection of nucleic acids that are present in atypical conformations is a crucial trigger of the innate immune response. Human Z-DNA binding protein 1 (ZBP1) is a pattern recognition receptor that harbors two Zα domains that recognize Z-DNA and Z-RNA. ZBP1 detects this alternate nucleic acid conformation as foreign, and upon stabilization of these substrates, it triggers activation of an immune response. Here, we present the backbone chemical shift assignment of a construct encompassing the Zα1 and Zα2 domains as well as the interconnecting linker of ZBP1. These assignments can be directly transferred to the isolated Zα1 and Zα2 domains, thereby demonstrating that these domains maintain virtually identical structures in the tandem context.

Rational development of functional hydrophilic polymer to characterize site-specific glycan differences between bovine milk and colostrum

As vastly modified on secreted proteins, N-glycosylation is found on milk proteins and undergo dynamic changes during lactation, characterizing milk protein glycosylation would benefit the elucidation of glycosylation pattern differences between samples. However, their low abundance required specific enrichment. Herein, through rational design and controllable synthesis, we developed a novel multi-functional polymer for the isolation of protein glycosylation. It efficiently separated glycopeptides from complex background inferences with mutual efforts of hydrophilic interaction chromatography (HILIC), metal ion affinity and ion exchange. By fine-tuning Ca2+ as regulators of aldehyde hyaluronic acid (HA) conformation, the grafting density of HA was remarkably improved. Moreover, grafting Ti4+ further enhanced the enrichment performance. Application of this material to characterize bovine milk and colostrum proteins yields 479 and 611 intact glycopeptides, respectively. Comparative analysis unraveled the distinct glycosylation pattern as well the different distribution of glycoprotein abundances between the two samples, offering insights for functional food development.

Mechanistic Insights into UV Spectral Changes of Pyruvic Acid and Pyruvate Part 1: Interaction with Water Molecules

We investigate how the UV spectra of pyruvic acid (PA) and pyruvate are impacted by interactions with water molecules. In particular, we would like to understand the mechanistic origin of the blue shift in the n →− π∗ transition. Pyruvic acid is the simplest α-keto organic acid and is common in the environment. We use density functional theory to optimize geometries to determine excitation energies and find that the excitation energies of the two main pyruvic acid conformers and pyruvate blue shift when interacting with 1 to 4 water molecules, both in vacuo and in a solvent. The excitation wavelength is blue-shifted by 0.9-9.2 nm when adding water molecules to the lowest energy conformer of PA. Calculations of the UV spectra of pyruvic acid (PA) and pyruvate are crucial for understanding the impact of the interactions with water molecules.

Computational Analysis of Albaflavenone Interaction with SlMAPK1 for Drought Resistance in Tomato.

As global agricultural challenges intensify, particularly drought stress, the exploration of innovative strategies for crop resilience has become crucial. This study focuses on the role of the microbial endophyte metabolite Albaflavenone in enhancing drought resistance in tomato (Solanum lycopersicum L.) through the activation of the SlMAPK1 protein in the MAPK pathway. To computationally analyze the interaction between Albaflavenone and SlMAPK1 and to elucidate the potential enhancement of drought tolerance in tomato plants through this interaction. We utilized molecular docking, homology modeling, and molecular dynamics simulations to investigate the binding affinities and interaction dynamics between SlMAPK1 and Albaflavenone. Functional network analysis was employed to examine protein-protein interactions within the MAPK pathway, while the MM-GBSA method was used to calculate binding free energies. Our computational analyses revealed that Albaflavenone exhibited a high binding affinity to SlMAPK1 with a binding energy of - 8.9kcal/mol. Molecular dynamics simulations showed this interaction significantly stabilized SlMAPK1, suggesting enhanced activity. Specifically, the root mean square deviation (RMSD) of the Albaflavenone-SlMAPK1 complex stabilized at around 3.1Å, while the root mean square fluctuations (RMSF) indicated consistent amino acid conformations. Additionally, the radius of gyration (Rg) analysis demonstrated minimal variance, suggesting a compact and stable protein-ligand complex. The significant binding affinity between Albaflavenone and SlMAPK1 highlights the potential of leveraging plant-microbe interactions in developing sustainable agricultural practices. These findings also demonstrate the effectiveness of computational methods in dissecting complex biological interactions, contributing to a deeper understanding of plant resilience strategies against environmental stresses.

Conformer-Specific Photoelectron Spectroscopy of Carbonic Acid: H2CO3.

Carbonic acid (H2CO3) is a fundamental species in biological, ecological, and astronomical systems. However, its spectroscopic characterization is incomplete because of its reactive nature. The photoionization (PI) and the photoion mass-selected threshold photoelectron (ms-TPE) spectra of H2CO3 were obtained by utilizing vacuum ultraviolet (VUV) synchrotron radiation and double imaging photoelectron photoion coincidence spectroscopy. Two carbonic acid conformers, namely, cis-cis and cis-trans, were identified. Experimental adiabatic ionization energies (AIEs) of cis-cis and cis-trans H2CO3 were determined to be 11.27 ± 0.02 and 11.18 ± 0.03 eV, and the cation enthalpies of formation could be derived as ΔfH°0K = 485 ± 2 and 482 ± 3 kJ mol-1, respectively. The cis-cis conformer shows intense peaks in the ms-TPES that are assigned to the C=O/C-OH stretching mode, while the cis-trans conformer exhibits a long progression to which two C=O/C-OH stretching modes contribute. The TPE spectra allow for the sensitive and conformer-selective detection of carbonic acid in terrestrial experiments to better understand astrochemical reactions.

Synthesis and Enzymatic Incorporation of a Dual-App Nucleotide Probe That Reports Antibiotics-Induced Conformational Change in the Bacterial Ribosomal Decoding Site RNA.

Natural nucleosides are nonfluorescent and do not have intrinsic labels that can be readily utilized for analyzing nucleic acid structure and recognition. In this regard, researchers typically use the so-called "one-label, one-technique" approach to study nucleic acids. However, we envisioned that a responsive dual-app nucleoside system that harnesses the power of two complementing biophysical techniques namely, fluorescence and 19F NMR, will allow the investigation of nucleic acid conformations more comprehensively than before. We recently introduced a nucleoside analogue by tagging trifluoromethyl-benzofuran at the C5 position of 2'-deoxyuridine, which serves as an excellent fluorescent and 19F NMR probe to study G-quadruplex and i-motif structures. Taking forward, here, we report the development of a ribonucleotide version of the dual-app probe to monitor antibiotics-induced conformational changes in RNA. The ribonucleotide analog is derived by conjugating trifluoromethyl-benzofuran at the C5 position of uridine (TFBF-UTP). The analog is efficiently incorporated by T7 RNA polymerase to produce functionalized RNA transcripts. Detailed photophysical and 19F NMR of the nucleoside and nucleotide incorporated into RNA oligonucleotides revealed that the analog is structurally minimally invasive and can be used for probing RNA conformations by fluorescence and 19F NMR techniques. Using the probe, we monitored and estimated aminoglycoside antibiotics binding to the bacterial ribosomal decoding site RNA (A-site, a very important RNA target). While 2-aminopurine, a famous fluorescent nucleic acid probe, fails to detect structurally similar aminoglycoside antibiotics binding to the A-site, our probe reports the binding of different aminoglycosides to the A-site. Taken together, our results demonstrate that TFBF-UTP is a very useful addition to the nucleic acid analysis toolbox and could be used to devise discovery platforms to identify new RNA binders of therapeutic potential.

From Functional Fatty Acids to Potent and Selective Natural-Product-Inspired Mimetics via Conformational Profiling.

Fatty acids play important signaling roles in biology, albeit typically lacking potency or selectivity, due to their substantial conformational flexibility. While being recognized as having properties of potentially great value as therapeutics, it is often the case that the functionally relevant conformation of the natural fatty acid is not known, thereby complicating efforts to develop natural-product-inspired ligands that have similar functional properties along with enhanced potency and selectivity profiles. In other words, without structural information associated with a particular functional relationship and the hopelessly unbiased conformational preferences of the endogenous ligand, one is molecularly ill-informed regarding the precise ligand-receptor interactions that play a role in driving the biological activity of interest. To address this problem, a molecular strategy to query the relevance of distinct subpopulations of fatty acid conformers has been established through "conformational profiling", a process whereby a unique collection of chiral and conformationally constrained fatty acids is employed to deconvolute beneficial structural features that impart natural-product-inspired function. Using oleic acid as an example because it is known to engage a variety of receptors, including GPR40, GPR120, and TLX, a 24-membered collection of mimetics was designed and synthesized. It was then demonstrated that this collection contained members that have enhanced potency and selectivity profiles, with some being clearly biased for engagement of the GPCRs GPR40 and GPR120 while others were identified as potent and selective modulators of the nuclear receptor TLX. A chemical synthesis strategy that exploited the power of modern technology for stereoselective synthesis was critical to achieving success, establishing a common sequence of bond-forming reactions to access a disparate collection of chiral mimetics, whose conformational preferences are impacted by the nature of stereodefined moieties differentially positioned about the C18 skeleton of the parent fatty acid. Overall, this study establishes a foundation to fuel future programs aimed at developing natural-product-inspired fatty acid mimetics as valuable tools in chemical biology and potential therapeutic leads.

Toward the IR Detection of Carbonic Acid: Absorption and Emission Spectra

With the recent radioastronomical detection of cis-trans-carbonic acid (H2CO3) in a molecular cloud toward the Galactic center, the more stable but currently unobserved cis-cis conformer is shown here to have strong IR features. While the higher-energy cis-trans-carbonic acid was detected at millimeter and centimeter wavelengths, owing to its larger dipole moment, the vibrational structure of cis-cis-carbonic acid is more amenable to its observation at micron wavelengths. Even so, both conformers have relatively large IR intensities, and some of these fall in regions not dominated by polycyclic aromatic hydrocarbons. Water features may inhibit observation near the 2.75 μm hydride stretches, but other vibrational fundamentals and even overtones in the 5.5–6.0 μm range may be discernible with JWST data. This work has employed high-level, accurately benchmarked quantum chemical anharmonic procedures to compute exceptionally accurate rotational spectroscopic data compared to experiment. Such performance implies that the IR absorption and even cascade emission spectral features computed in this work should be accurate and will provide the needed reference for observation of either carbonic acid conformer in various astronomical environments.

Unraveling the complexity: Advanced methods in analyzing DNA, RNA, and protein interactions.

Exploring the intricate interplay within and between nucleic acids, as well as their interactions with proteins, holds pivotal significance in unraveling the molecular complexities steering cancer initiation and progression. To investigate these interactions, a diverse array of highly specific and sensitive molecular techniques has been developed. The selection of a particular technique depends on the specific nature of the interactions. Typically, researchers employ an amalgamation of these different techniques to obtain a comprehensive and holistic understanding of inter- and intramolecular interactions involving DNA-DNA, RNA-RNA, DNA-RNA, or protein-DNA/RNA. Examining nucleic acid conformation reveals alternative secondary structures beyond conventional ones that have implications for cancer pathways. Mutational hotspots in cancer often lie within sequences prone to adopting these alternative structures, highlighting the importance of investigating intra-genomic and intra-transcriptomic interactions, especially in the context of mutations, to deepen our understanding of oncology. Beyond these intramolecular interactions, the interplay between DNA and RNA leads to formations like DNA:RNA hybrids (known as R-loops) or even DNA:DNA:RNA triplex structures, both influencing biological processes that ultimately impact cancer. Protein-nucleic acid interactions are intrinsic cellular phenomena crucial in both normal and pathological conditions. In particular, genetic mutations or single amino acid variations can alter a protein's structure, function, and binding affinity, thus influencing cancer progression. It is thus, imperative to understand the differences between wild-type (WT) and mutated (MT) genes, transcripts, and proteins. The review aims to summarize the frequently employed methods and techniques for investigating interactions involving nucleic acids and proteins, highlighting recent advancements and diverse adaptations of each technique.

Mechanistic Insight into Poly-Reactivity of Immune Antibodies upon Acid Denaturation or Arginine Mutation in Antigen-Binding Regions.

The poly-reactivity of antibodies is defined as their binding to specific antigens as well as to related proteins and also to unrelated targets. Poly-reactivity can occur in individual molecules of natural serum antibodies, likely due to their conformation flexibility, and, for therapeutic antibodies, it plays a critical role in their clinical development. On the one hand, it can enhance their binding to target antigens and cognate receptors, but, on the other hand, it may lead to a loss of antibody function by binding to off-target proteins. Notably, poly-reactivity has been observed in antibodies subjected to treatments with dissociating, destabilizing or denaturing agents, in particular acidic pH, a common step in the therapeutic antibody production process involving the elution of Protein-A bound antibodies and viral clearance using low pH buffers. Additionally, poly-reactivity can emerge during the affinity maturation in the immune system, such as the germinal center. This review delves into the underlying potential causes of poly-reactivity, highlighting the importance of conformational flexibility, which can be further augmented by the acid denaturation of antibodies and the introduction of arginine mutations into the complementary regions of antibody-variable domains. The focus is placed on a particular antibody's acid conformation, meticulously characterized through circular dichroism, differential scanning calorimetry, and sedimentation velocity analyses. By gaining a deeper understanding of these mechanisms, we aim to shed light on the complexities of antibody poly-reactivity and its implications for therapeutic applications.

Machine learning models based on residue interaction network for ABCG2 transportable compounds recognition

As the one of the most important protein of placental transport of environmental substances, the identification of ABCG2 transport molecules is the key step for assessing the risk of placental exposure to environmental chemicals. Here, residue interaction network (RIN) was used to explore the difference of ABCG2 binding conformations between transportable and non-transportable compounds. The RIN were treated as a kind of special quantitative data of protein conformation, which not only reflected the changes of single amino acid conformation in protein, but also indicated the changes of distance and action type between amino acids. Based on the quantitative RIN, four machine learning algorithms were applied to establish the classification and recognition model for 1100 compounds with transported by ABCG2 potential. The random forest (RF) models constructed with RIN presented the best and satisfied predictive ability with an accuracy of training set of 0.97 and the test set of 0.96 respectively. In conclusion, the construction of residue interaction network provided a new perspective for the quantitative characterization of protein conformation and the establishment of prediction models for transporter molecular recognition. The ABCG2 transport molecular recognition model based on residue interaction network provides a possible way for screening environmental chemistry transported through placenta.

Molecular Dynamics of Artificially Pair-Decoupled Systems: An Accurate Tool for Investigating the Importance of Intramolecular Couplings.

We propose a numerical technique to accurately simulate the vibrations of organic molecules in the gas phase, when pairs of atoms (or, in general, groups of degrees of freedom) are artificially decoupled, so that their motion is instantaneously decorrelated. The numerical technique we have developed is a symplectic integration algorithm that never requires computation of the force but requires estimates of the Hessian matrix. The theory we present to support our technique postulates a pair-decoupling Hamiltonian function, which parametrically depends on a decoupling coefficient α ∈ [0, 1]. The closer α is to 0, the more decoupled the selected atoms. We test the correctness of our numerical method on small molecular systems, and we apply it to study the vibrational spectroscopic features of salicylic acid at the Density Functional Theory ab initio level on a fitted potential. Our pair-decoupled simulations of salicylic acid show that decoupling hydrogen-bonded atoms do not significantly influence the frequencies of stretching modes, but enhance enormously the out-of-plane wagging and twisting motions of the hydroxyl and carboxyl groups to the point that the carboxyl and hydroxyl groups may overcome high potential energy barriers and change the salicylic acid conformation after a short simulation time. In addition, we found that the acidity of salicylic acid is more influenced by the dynamical couplings of the proton of the carboxylic group with the carbon ring than with the hydroxyl group.

Single selenium atom to control nucleic acid conformation and large crystal growth

Single selenium atom to control nucleic acid conformation and large crystal growth

Conformational State of Fenamates at the Membrane Interface: A MAS NOESY Study.

The present work analyzes the 1H NOESY MAS NMR spectra of three fenamates (mefenamic, tolfenamic, and flufenamic acids) localized in the lipid-water interface of phosphatidyloleoylphosphatidylcholine (POPC) membranes. The observed cross-peaks in the two-dimensional NMR spectra characterized intramolecular proximities between the hydrogen atoms of the fenamates as well as intermolecular interactions between the fenamates and POPC molecules. The peak amplitude normalization for an improved cross-relaxation (PANIC) approach, the isolated spin-pair approximation (ISPA) model, and the two-position exchange model were used to calculate the interproton distances indicative of specific conformations of the fenamates. The results showed that the proportions of the A+C and B+D conformer groups of mefenamic and tolfenamic acids in the presence of POPC were comparable within the experimental error and amounted to 47.8%/52.2% and 47.7%/52.3%, respectively. In contrast, these proportions for the flufenamic acid conformers differed and amounted to 56.6%/43.4%. This allowed us to conclude that when they bind to the POPC model lipid membrane, fenamate molecules change their conformational equilibria.

Hygienic pelleting can decrease Hubbard × Ross 708 apparent ileal amino acid digestibility, broiler performance, and increase digestible amino acid requirement

To reduce pathogenic bacteria such as Salmonella in feed and ensure a high-quality pellet, diets for broilers, turkeys, and breeders may be pelleted using high conditioning temperatures and extended times. However, increased conditioning temperature and time may alter amino acid conformation regarding secondary and tertiary structure, decrease digestibility and subsequent bird performance, and necessitate increased amino acid formulation density. The objective of this study was to evaluate increased conditioning temperature and exposure time within the pelleting process using a hygienizer (hygienic pelleting) in diets that vary in digestible lysine and corresponding amino acid ratio on Hubbard × Ross 708 starter broiler performance, apparent ileal amino acid digestibility (AIAAD), and requirement. A 5 × 2 factorial arrangement of treatments with 5 levels of digestible lysine (−20, −10%, 2019 Ross Broiler Starter Nutrition recommendation, +10, and +20%) and 2 techniques of pelleting (standard and hygienic) was utilized in a randomized complete block design. Standard pelleting utilized 77°C conditioning for 30 s and hygienic pelleting utilized 88°C conditioning for 60 s and 6-min retention time in a hygienizer. Each treatment was fed to 12 replicate pens of 10 male broiler Hubbard × Ross 708 chicks for a 21-day period. Standard pelleting and hygienic pelleting demonstrated a 1.19 and 1.37% digestible Lys requirement, respectively, using the 2 straight line, 1-breakpoint broken-line regression model. Performance data, AIAAD, and the broken-line regression model suggest an approximate 10% increase in digestible Lys and corresponding amino acid ratio relative to 2019 Ross Broiler Starter Nutrition Specifications for Hygienic Pelleting. Nutritionists should be cognizant that nutritional detriment may accompany procedures used to create hygienic pelleted feed.

Spectral extension of two glycolic acid conformers into the submillimeter regime

The rotational spectrum of glycolic acid has been extended to frequencies from 318 to 1000 GHz. Analysis of this spectrum builds upon previous experimental and theoretical studies performed on glycolic acid conformers. Extensive new assignments were made for the ground vibrational state of the SSC conformer of glycolic acid in addition to the two lowest vibrational states. Assignments were also expanded for the AAT conformer of glycolic acid. This work improves the precision of some rotational constants and nearly all of the centrifugal distortion constants for each conformer and vibrational state. The spectral predictions based on these refined parameters will facilitate the search for glycolic acid in the interstellar medium at frequencies covered by submillimeter observatories.

Exploring the Conformational Equilibrium of Mefenamic Acid Released from Silica Aerogels via NMR Analysis.

This study examines the influence of mefenamic acid on the physical and chemical properties of silica aerogels, as well as its effect on the sorption characteristics of the composite material. Solid state magic angle spinning nuclear magnetic resonance (MAS NMR) and high-pressure 13C NMR kinetic studies were conducted to identify the presence of mefenamic acid and measure the kinetic rates of CO2 sorption. Additionally, a high-pressure T1-T2 relaxation-relaxation correlation spectroscopy (RRCOSY) study was conducted to estimate the relative amount of mefenamic acid in the aerogel's pores, and a high-pressure nuclear Overhauser effect spectoscopy (NOESY) study was conducted to investigate the conformational preference of mefenamic acid released from the aerogel. The results indicate that mefenamic acid is affected by the chemical environment of the aerogel, altering the ratio of mefenamic acid conformers from 75% to 25% in its absence to 22% to 78% in the presence of aerogel.

Cavity hairpin ThT-light nucleic acid switches: the construction of label- and enzyme-free sensing and imaging platforms.

Thioflavin T (ThT) is a classical fluorescent dye gaining prominence in current research regarding nucleic acid conformations (NACs). However, most NACs with the ability to excite ThT fluorescent are unique or form in demanding conditions, limiting the extensiveness and depth of ThT application in sensing and imaging. Therefore, this study proposed CGG-AAA mismatched cavity hairpin ThT-light nucleic acid switches (CHTLNAS) with excellent fluorescence excitation over 500-fold higher than spontaneous, 17∼20-fold higher than ssDNA and 2.5∼5-fold higher than complementary duplex. Based on the excellent fluorescence excitation, convenient conformation formation, good sequence programmability, and flexible allosteric ability (known as the Worm-crack pod mechanism mediated by the target), it achieved the label- and enzyme-free detection of tetracycline (TET) and berberine (BB) at the pM level within 10 min. Moreover, it was found enable to realize the sensitive tracking of intracellular carriers at the nM level of ThT entry concentration, and prolongated its cell nuclear-entry time of ThT over 8 h, overcoming the non-specific high background signal interference of ThT in the nuclear region, and expanding the diversified application of ThT in cell biology research. Therefore, CHTLNAS is a more universal, practical tool than G-quadruplex or other kinds of NACs for ThT development and utilization in sensing and imaging platforms.