Dynamics Of Structural Transitions Research Articles

Oligo-Adenine Derived Secondary Nucleic Acid Frameworks: From Structural Characteristics to Applications.

Oligo-adenine (polyA) is primarily known for its critical role in mRNA stability, translational status, and gene regulation. Beyond its biological functions, extensive research has unveiled the diverse applications of polyA. In response to environmental stimuli, single polyA strands undergo distinctive structural transitions into diverse secondary configurations, which are reversible upon the introduction of appropriate counter-triggers. In this review, we systematically summarize recent advances of noncanonical structures derived from polyA, including A-motif duplex, A-cyanuric acid triplex, A-coralyne-A duplex, and T ⋅ A-T triplex. The structural characteristics and mechanisms underlying these conformations under specific external stimuli are addressed, followed by examples of their applications in stimuli-responsive DNA hydrogels, supramolecular fibre assembly, molecular electronics and switches, biosensing and bioengineering, payloads encapsulation and release, and others. A detailed comparison of these polyA-derived noncanonical structures is provided, highlighting their distinctive features. Furthermore, by integrating their stimuli-responsiveness and conformational characteristics, advanced material development, such as pH-cascaded DNA hydrogels and supramolecular fibres exhibiting dynamic structural transitions adapting environmental cues, are introduced. An outlook for future developments is also discussed. These polyA derived, stimuli-responsive, noncanonical structures enrich the arsenal of DNA "toolbox", offering dynamic DNA frameworks for diverse future applications.

Dynamic three-dimensional structures of a metal–organic framework captured with femtosecond serial crystallography

Crystalline systems consisting of small-molecule building blocks have emerged as promising materials with diverse applications. It is of great importance to characterize not only their static structures but also the conversion of their structures in response to external stimuli. Femtosecond time-resolved crystallography has the potential to probe the real-time dynamics of structural transitions, but, thus far, this has not been realized for chemical reactions in non-biological crystals. In this study, we applied time-resolved serial femtosecond crystallography (TR-SFX), a powerful technique for visualizing protein structural dynamics, to a metal–organic framework, consisting of Fe porphyrins and hexazirconium nodes, and elucidated its structural dynamics. The time-resolved electron density maps derived from the TR-SFX data unveil trifurcating structural pathways: coherent oscillatory movements of Zr and Fe atoms, a transient structure with the Fe porphyrins and Zr6 nodes undergoing doming and disordering movements, respectively, and a vibrationally hot structure with isotropic structural disorder. These findings demonstrate the feasibility of using TR-SFX to study chemical systems.

Dynamic visible-infrared transmittance regulation based on fluoroaluminate glass by femtosecond-laser-customized induced microstructure

Herein, we report significant microstructure-induced dynamic visible–infrared transmittance regulation on a novel fluoroaluminate glass (AlF3) via femtosecond-laser-customized fabrication technology. Fluoroaluminate glass, which has excellent optical transmittance in the visible–infrared wavelength range (400–5000 nm), is a candidate for visible and infrared optical devices. Based on its performance, three types of microstructures or material transitions (Snow-Layer Microstructure (SLS), Surface-Ablation Microstructure (SAS), and Interior-Modified Microstructure (IMS)) were successfully achieved using a femtosecond-laser-customized microstructure, including surface and interior fabrication routes with cylindrical lens focusing. Dynamic and reversible structural transition features were observed between the snow-layer and surface-ablation microstructures. These types of structures showed different transmittance regulation trends at visible and infrared wavelengths. The transmittance is decreased for IMS, SAS, and SLS in the visible region, while in the infrared region the transmittance increased for IMS and decreased for SAS and SLS. Based on Raman spectroscopy analyses, the chemical structures of the SAS before and after femtosecond-laser ablation appeared to remain constant, whereas the SLS showed a significant difference, giving rise to different transmittance regulations. This study provides a novel fabrication method and strategy for regulating visible and infrared transmission and promotes smart application of glass materials.

The ubiquitination landscape of the influenza A virus polymerase

During influenza A virus (IAV) infections, viral proteins are targeted by cellular E3 ligases for modification with ubiquitin. Here, we decipher and functionally explore the ubiquitination landscape of the IAV polymerase proteins during infection of human alveolar epithelial cells by applying mass spectrometry analysis of immuno-purified K-ε-GG (di-glycyl)-remnant-bearing peptides. We have identified 59 modified lysines across the three subunits, PB2, PB1 and PA of the viral polymerase of which 17 distinctively affect mRNA transcription, vRNA replication and the generation of recombinant viruses via non-proteolytic mechanisms. Moreover, further functional and in silico analysis indicate that ubiquitination at K578 in the PB1 thumb domain is mechanistically linked to dynamic structural transitions of the viral polymerase that are required for vRNA replication. Mutations K578A and K578R differentially affect the generation of recombinant viruses by impeding cRNA and vRNA synthesis, NP binding as well as polymerase dimerization. Collectively, our results demonstrate that the ubiquitin-mediated charge neutralization at PB1-K578 disrupts the interaction to an unstructured loop in the PB2 N-terminus that is required to coordinate polymerase dimerization and facilitate vRNA replication. This provides evidence that IAV exploits the cellular ubiquitin system to modulate the activity of the viral polymerase for viral replication.

Guidelines for the Simulations of Nitroxide X-Band cw EPR Spectra from Site-Directed Spin Labeling Experiments Using SimLabel.

Site-directed spin labeling (SDSL) combined with continuous wave electron paramagnetic resonance (cw EPR) spectroscopy is a powerful technique to reveal, at the local level, the dynamics of structural transitions in proteins. Here, we consider SDSL-EPR based on the selective grafting of a nitroxide on the protein under study, followed by X-band cw EPR analysis. To extract valuable quantitative information from SDSL-EPR spectra and thus give a reliable interpretation on biological system dynamics, a numerical simulation of the spectra is required. However, regardless of the numerical tool chosen to perform such simulations, the number of parameters is often too high to provide unambiguous results. In this study, we have chosen SimLabel to perform such simulations. SimLabel is a graphical user interface (GUI) of Matlab, using some functions of Easyspin. An exhaustive review of the parameters used in this GUI has enabled to define the adjustable parameters during the simulation fitting and to fix the others prior to the simulation fitting. Among them, some are set once and for all (gy, gz) and others are determined (Az, gx) thanks to a supplementary X-band spectrum recorded on a frozen solution. Finally, we propose guidelines to perform the simulation of X-band cw-EPR spectra of nitroxide labeled proteins at room temperature, with no need of uncommon higher frequency spectrometry and with the minimal number of variable parameters.

Orbital-Hybridization-Driven Charge Density Wave Transition in CsV3 Sb5 Kagome Superconductor.

Owing to its inherent non-trivial geometry, the unique structural motif of the recently discovered kagome topological superconductor AV3 Sb5 (A = K, Rb, Cs) is an ideal host of diverse topologically non-trivial phenomena, including giant anomalous Hall conductivity, topological charge order, charge density wave (CDW), and unconventional superconductivity. Despite possessing a normal-state CDW order in the form of topological chiral charge order and diverse superconducting gaps structures, it remains unclear how fundamental atomic-level properties and many-body effects including Fermi surface nesting, electron-phonon coupling, and orbital hybridization contribute to these symmetry-breaking phenomena. Here, the direct participation of the V3d-Sb5p orbital hybridization in mediating the CDW phase transition in CsV3 Sb5 is reported. The combination of temperature-dependent X-ray absorption and first-principles studies clearly indicates the inverse Star-of-David structure as the preferred reconstruction in the low-temperature CDW phase. The results highlight the critical role that Sb orbitals play and establish orbital hybridization as the direct mediator of the CDW states and structural transition dynamics in kagome unconventional superconductors. This is a significant step toward the fundamental understanding and control of the emerging correlated phases from the kagome lattice through the orbital interactions and provides promising approaches to novel regimes in unconventional orders and topology.

Tripodal Quinone-cyanine Fluorescent Dye as a Tool for Analyzing Structural Dynamics of DNA Guanine-quadruplex

Tripodal quinone-cyanine fluorescent dye, QCy(MeBT)3, which responds to DNA double-strand and guanine-quadruplex (G4) with different wavelength fluorescence emissions, was applied for analyzing the structural transition dynamics between DNA double-strand and G4. Results indicated that the formation and disruption of DNA double-strand and G4 could be monitored with 700 nm and 600 nm fluorescence. As QCy(MeBT)3 can monitor not only G4 formation but also G4 disruption, the system might contribute largely to the screening of both G4 ligands and G4 disruptors.

Microfluidic Nanomaterial Synthesis and In Situ SAXS, WAXS, or SANS Characterization: Manipulation of Size Characteristics and Online Elucidation of Dynamic Structural Transitions.

With the ability to cross biological barriers, encapsulate and efficiently deliver drugs and nucleic acid therapeutics, and protect the loaded cargos from degradation, different soft polymer and lipid nanoparticles (including liposomes, cubosomes, and hexosomes) have received considerable interest in the last three decades as versatile platforms for drug delivery applications and for the design of vaccines. Hard nanocrystals (including gold nanoparticles and quantum dots) are also attractive for use in various biomedical applications. Here, microfluidics provides unique opportunities for the continuous synthesis of these hard and soft nanomaterials with controllable shapes and sizes, and their in situ characterization through manipulation of the flow conditions and coupling to synchrotron small-angle X-ray (SAXS), wide-angle scattering (WAXS), or neutron (SANS) scattering techniques, respectively. Two-dimensional (2D) and three-dimensional (3D) microfluidic devices are attractive not only for the continuous production of monodispersed nanomaterials, but also for improving our understanding of the involved nucleation and growth mechanisms during the formation of hard nanocrystals under confined geometry conditions. They allow further gaining insight into the involved dynamic structural transitions, mechanisms, and kinetics during the generation of self-assembled nanostructures (including drug nanocarriers) at different reaction times (ranging from fractions of seconds to minutes). This review provides an overview of recently developed 2D and 3D microfluidic platforms for the continuous production of nanomaterials, and their simultaneous use in in situ characterization investigations through coupling to nanostructural characterization techniques (e.g., SAXS, WAXS, and SANS).

Direct observation of protein structural transitions through entire amyloid aggregation processes in water using 2D-IR spectroscopy.

Amyloid proteins that undergo self-assembly to form insoluble fibrillar aggregates have attracted much attention due to their role in biological and pathological significance in amyloidosis. This study aims to understand the amyloid aggregation dynamics of insulin (INS) in H2O using two-dimensional infrared (2D-IR) spectroscopy. Conventional IR studies have been performed in D2O to avoid spectral congestion despite distinct H–D isotope effects. We observed a slowdown of the INS fibrillation process in D2O compared to that in H2O. The 2D-IR results reveal that different quaternary structures of INS at the onset of the nucleation phase caused the distinct fibrillation pathways of INS in H2O and D2O. A few different biophysical analysis, including solution-phase small-angle X-ray scattering combined with molecular dynamics simulations and other spectroscopic techniques, support our 2D-IR investigation results, providing insight into mechanistic details of distinct structural transition dynamics of INS in water. We found the delayed structural transition in D2O is due to the kinetic isotope effect at an early stage of fibrillation of INS in D2O, i.e., enhanced dimer formation of INS in D2O. Our 2D-IR and biophysical analysis provide insight into mechanistic details of structural transition dynamics of INS in water. This study demonstrates an innovative 2D-IR approach for studying protein dynamics in H2O, which will open the way for observing protein dynamics under biological conditions without IR spectroscopic interference by water vibrations.

Dynamic Structural Changes and Thermodynamics in Phase Separation Processes of an Intrinsically Disordered-Ordered Protein Model.

Elastin‐like proteins (ELPs) are biologically important proteins and models for intrinsically disordered proteins (IDPs) and dynamic structural transitions associated with coacervates and liquid–liquid phase transitions. However, the conformational status below and above coacervation temperature and its role in the phase separation process is still elusive. Employing matrix least‐squares global Boltzmann fitting of the circular dichroism spectra of the ELPs (VPGVG)20, (VPGVG)40, and (VPGVG)60, we found that coacervation occurs sharply when a certain number of repeat units has acquired β‐turn conformation (in our sequence setting a threshold of approx. 20 repeat units). The character of the differential scattering of the coacervate suspensions indicated that this fraction of β‐turn structure is still retained after polypeptide assembly. Such conformational thresholds may also have a role in other protein assembly processes with implications for the design of protein‐based smart materials.

Decoding the Structural Dynamics and Conformational Alternations of DNA Secondary Structures by Single-Molecule FRET Microspectroscopy.

In addition to the canonical double helix form, DNA is known to be extrapolated into several other secondary structural patterns involving themselves in inter- and intramolecular type hydrogen bonding. The secondary structures of nucleic acids go through several stages of multiple, complex, and interconvertible heterogeneous conformations. The journey of DNA through these conformers has significant importance and has been monitored thoroughly to establish qualitative and quantitative information about the transition between the unfolded, folded, misfolded, and partially folded states. During this structural interconversion, there always exist specific populations of intermediates, which are short-lived or sometimes even do not accumulate within a heterogeneous population and are challenging to characterize using conventional ensemble techniques. The single-molecule FRET(sm-FRET) microspectroscopic method has the advantages to overcome these limitations and monitors biological phenomena transpiring at a measurable high rate and balanced stochastically over time. Thus, tracing the time trajectory of a particular molecule enables direct measurement of the rate constant of each transition step, including the intermediates that are hidden in the ensemble level due to their low concentrations. This review is focused on the advantages of the employment of single-molecule Forster’s resonance energy transfer (sm-FRET), which is worthwhile to access the dynamic architecture and structural transition of various secondary structures that DNA adopts, without letting the donor of one molecule to cross-talk with the acceptor of any other. We have emphasized the studies performed to explore the states of folding and unfolding of several nucleic acid secondary structures, for example, the DNA hairpin, Holliday junction, G-quadruplex, and i-motif.

Structural insights into the dynamic and controlled multiphase evolution of layered-spinel heterostructured sodium oxide cathode

Concepts in sodium oxide cathode composite structures have captured widespread attention; however, the detailed monitoring and accurate control of composite structural evolution during charge and discharge process remain challenging, especially for complex triphases and other similar systems. Here, we report the accurate manipulation of multiphase structural evolution during high-voltage cycling with improved electrochemical performance and adjustable sodium ion intercalation/deintercalation electrochemical behavior via rational chemical element substitution using a series of Fd -3 m spinel and layered P2/P3 heterostructures as proof-of-concept materials. Multiphase evolutions as a function of chemical substitution during high-voltage cycling are demonstrated. Meanwhile, we also monitor the dynamic formation process during calcination and observe the atomic arrangement of triphase heterostructure through various advanced characterization techniques. Overall, this study reveals controllable multiphase structural evolution in a model system and explores the related fundamental science required for future development of high-performance sodium-ion batteries. A series of spinel and P2/P3 heterostructures are studied Dynamic structural evolution and controllable multiphase transition are confirmed The architecture of controlled multiphase structural evolution is demonstrated Xiao et al. demonstrate the accurate manipulation of multiphase structural evolution during high-voltage cycling using a series of Fd-3 m spinel and layered P2/P3 heterostructures as proof-of-concept materials. The concept of controllable multiphase structural evolution architecture in a model system is realized, which may have implications for design of high-performance sodium-ion batteries.

A Revisit to Turnover Kinetics of Individual Escherichia coli β-Galactosidase Molecules.

Single-molecule experiments on β-galactosidase from Escherichia coli that catalyzes the hydrolysis of resorufin-β-d-galactopyranoside revealed important observations like fluctuating catalytic rate, memory effects arising from temporal correlations between the enzymatic turnovers and nonexponential waiting time distributions. The root cause of the observed results is intrinsic fluctuations among the different conformers of the active species, during the course of the reaction, thereby imparting dynamic disorder in the system under investigation. Originally, a multistate stochastic kinetic theory was employed that, despite satisfying the measured waiting time distributions and the mean waiting times at different substrate concentrations, yields a constant estimate of the randomness parameter. Inevitably, this manifests a strong disagreement with the substrate-concentration-dependent time variations of the said distribution, which at the same time misinterprets the measured magnitudes of the randomness parameter at lower concentrations. Here, we suggest a dual approach to the single-enzyme reaction, independently, making important improvements over the parent study and the recently suggested two-state stochastic analyses followed by quantitative rationalization of the experimental data. In the first case, an off-pathway mechanism satisfied the Michaelis-Menten equation under the circumstance of prevailing disorder while tested against the single-molecule data. However, recovery of randomness data in the lower-concentration regime, albeit primarily marks a significant refinement, a qualitative agreement at the growing concentrations seems to be reasoned by an account of switching among the limited numbers of discrete conformers. Consequently, in the second case, we circumvented the conventional way of approaching the enzyme catalysis and mapped the dynamics of structural transitions of the biocatalyst with the temporal fluctuations of the spatial distance between the different locations along a coarse-grained polymer chain. Exploiting a general mechanism for dynamic disorder, a reaction-diffusion formalism yielded an analytical expression for the waiting time distribution of the enzymatic turnovers, from which the mean waiting time and the randomness parameter were readily determined. Application of our results to the findings of the experiment on single β-galactosidase shows a quantitative agreement in each case. This soundly validates the usefulness of accounting for a more rigorous microscopic description pertinent to the conformational multiplicity in rationalizing the real-time data over the routine state-based sketch of the reaction system.

Spatially and time-resolved SAXS for monitoring dynamic structural transitions during in situ generation of non-lamellar liquid crystalline phases in biologically relevant media

Formation of high viscous inverse lyotropic liquid crystalline phases in situ upon exposure of low viscous drug-loaded lipid preformulations to synovial fluid provides a promising approach for design of depot formulations for intra-articular drug delivery. Rational formulation design relies on a fundamental understanding of the synovial fluid-mediated dynamic structural transitions occurring at the administration site. At conditions mimicking the in vivo situation, we investigated in real-time such transitions at multiple positions by synchrotron small-angle X-ray scattering (SAXS) combined with an injection-cell. An injectable diclofenac-loaded quaternary preformulation consisting of 72/8/10/10% (w/w) glycerol monooleate/1,2-dioleoyl-glycero-3-phospho-rac-(1-glycerol)/ethanol/water was injected into hyaluronic acid solution or synovial fluid. A fast generation of a coherent drug depot of inverse bicontinuous Im3m and Pn3m cubic phases was observed. Through construction of 2D spatial maps from measurements performed 60 min after injection of the preformulation, it was possible to differentiate liquid crystalline rich- and excess hyaluronic acid solution- or synovial fluid-rich regimes. Synchrotron SAXS findings confirmed that the exposure of the preformulation to the media leads to alterations in structural features in position- and time-dependent manners. Effects of biologically relevant medium composition on the structural features, and implications for development of formulations with sustained drug release properties are highlighted.

A General Fret-Based Method to Monitor Protein Oligomerization and Condensate Phase Transition in Cells

Proteins can self-assemble into a wide range of architectures, ranging from microscopically invisible small oligomers to condensates of varying structure and dynamics. These assemblies are essential for the functioning of the cell and are associated with many kinds of diseases. Because the intracellular environment influences assemblies concerning stoichiometry, structure, and dynamics, which is due to chaperone action or nonspecific interactions, it pertinent that these assemblies are monitored in a minimally invasive manner inside cells. Here we present a FRET-based method that allows detailed monitoring of protein self-assembly. The sensitive nature of this method allows monitoring the dynamic aggregation behavior and distinguishing different types of assemblies. We thus observe the dynamic structural transition between unstructured condensates and fibrils and confirm an array of previous observations on aggregations in yeast and HEK293. In addition, we find a distinct cell-to-cell variation in kinetics and structure of the respective condensates. This method is modular and easily implemented in other native or non-native proteins and will provide crucial insight into protein self-assembly.

Structural Transition Dynamics in Carbon Electrode‐Based Single‐Molecule Junctions

AbstractMonitoring the structural transitions of individual molecules is of great significance because it helps to in depth explore the properties of molecules and provide diverse possibilities for molecular applications in the fields of chemistry, biology and material science. This review summarizes the strategy of using single‐molecule electrical approaches to study molecular structure transitions at the single‐molecule level in real time. Specifically, through the use of stable single‐molecule devices for real‐time electrical monitoring, the process of molecular structure transitions of a single molecule can be investigated, which helps to explore the nature of molecules in chemical and biological systems. In particular, the detection methods have been extended to the investigation of biological macromolecules for monitoring the conformational changes of nucleotide chains in different systems, such as double helix DNA, aptamer and DNA enzyme. In the end, we discuss the future challenges of probing structural transitions of single molecules, and provide prospects for further breakthroughs in this field.What is the most favorite and original chemistry developed in your research group?Single‐molecule device fabrication and detection.How do you get into this specific field? Could you please share some experiences with our readers?I started to pursue this field during my PhD period. The promise of molecular electronics that might breakthrough the development bottleneck of current microelectronics inspired my great interest. This interest drives me to persist in the pursuit of practical molecular electronic devices as perfect as possible.What is the most important personality for scientific research?Three aspects: interest, persistence and industriousness. The first key to the success should be diligence.How do you keep balance between research and family?It is hard to balance the time. It is very important to have understanding and support from the family. I love them.What's your hobbies?Driving and watching movies.

In situ observation of the crystal structure transition of Pt-Sn intermetallic nanoparticles during deactivation and regeneration.

The mechanism of the deactivation and regeneration of PtSn intermetallic compound nanoparticle (iNP) catalysts was studied by in situ TEM investigation. Our study reveals the reversible dynamic structural transition of the iNPs during deactivation and regeneration, which provides a direct correlation between the atomic structure and the catalytic activity of the iNPs.

Mechanistic insights into the synergistic activation of the RXR–PXR heterodimer by endocrine disruptor mixtures

Humans are chronically exposed to mixtures of xenobiotics referred to as endocrine-disrupting chemicals (EDCs). A vast body of literature links exposure to these chemicals with increased incidences of reproductive, metabolic, or neurological disorders. Moreover, recent data demonstrate that, when used in combination, chemicals have outcomes that cannot be predicted from their individual behavior. In its heterodimeric form with the retinoid X receptor (RXR), the pregnane X receptor (PXR) plays an essential role in controlling the mammalian xenobiotic response and mediates both beneficial and detrimental effects. Our previous work shed light on a mechanism by which a binary mixture of xenobiotics activates PXR in a synergistic fashion. Structural analysis revealed that mutual stabilization of the compounds within the ligand-binding pocket of PXR accounts for the enhancement of their binding affinity. In order to identify and characterize additional active mixtures, we combined a set of cell-based, biophysical, structural, and in vivo approaches. Our study reveals features that confirm the binding promiscuity of this receptor and its ability to accommodate bipartite ligands. We reveal previously unidentified binding mechanisms involving dynamic structural transitions and covalent coupling and report four binary mixtures eliciting graded synergistic activities. Last, we demonstrate that the robust activity obtained with two synergizing PXR ligands can be enhanced further in the presence of RXR environmental ligands. Our study reveals insights as to how low-dose EDC mixtures may alter physiology through interaction with RXR-PXR and potentially several other nuclear receptor heterodimers.

The Conformational Equilibrium of the Neuropeptide Y2 Receptor in Bilayer Membranes.

Dynamic structural transitions within the seven‐transmembrane bundle represent the mechanism by which G‐protein‐coupled receptors convert an extracellular chemical signal into an intracellular biological function. Here, the conformational dynamics of the neuropeptide Y receptor type 2 (Y2R) during activation was investigated. The apo, full agonist‐, and arrestin‐bound states of Y2R were prepared by cell‐free expression, functional refolding, and reconstitution into lipid membranes. To study conformational transitions between these states, all six tryptophans of Y2R were 13C‐labeled. NMR‐signal assignment was achieved by dynamic‐nuclear‐polarization enhancement and the individual functional states of the receptor were characterized by monitoring 13C NMR chemical shifts. Activation of Y2R is mediated by molecular switches involving the toggle switch residue Trp2816.48 of the highly conserved SWLP motif and Trp3277.55 adjacent to the NPxxY motif. Furthermore, a conformationally preserved “cysteine lock”‐Trp11623.50 was identified.

In situ monitoring of the formation of lipidic non-lamellar liquid crystalline depot formulations in synovial fluid

Administration of parenteral liquid crystalline phases, forming in-vivo with tunable nanostructural features and sustained release properties, offers an attractive approach for treatment of infections and local drug delivery. It has also a potential use for postoperative pain management after arthroscopic knee surgery. However, the optimal use of this drug delivery principle requires an improved understanding of the involved dynamic structural transitions after administration of low-viscous stimulus-responsive lipid precursors and their fate after direct contact with the biological environment. These precursors (preformulations) are typically based on a single biologically relevant lipid (or a lipid combination) with non-lamellar liquid crystalline phase forming propensity. In relation to liquid crystalline depot design for intra-articular drug delivery, it was our interest in the present study to shed light on such dynamic structural transitions by combining synchrotron SAXS with a remote controlled addition of synovial fluid (or buffer containing 2% (w/v) albumin). This combination allowed for monitoring in real-time the hydration-triggered dynamic structural events on exposure of the lipid precursor (organic stock solution consisting of the binary lipid mixture of monoolein and castor oil) to excess synovial fluid (or excess buffer). The synchrotron SAXS findings indicate a fast generation of inverse bicontinuous cubic phases within few seconds. The effects of (i) the organic solvent N-methyl-2-pyrolidone (NMP), (ii) the lipid composition, and (iii) the albumin content on modulating the structures of the self-assembled lipid aggregates and the implications of the experimental findings in the design of liquid crystalline depots for intra-articular drug delivery are discussed.