Folding Process Research Articles

Investigation on the Torsional–Flexural Instability Phenomena during the Bending Process of Hairpin Windings: Experimental Tests and FE Model Validation

Modern electric motors developed for the automotive industry have an ever higher power density with a relatively compact size. Among the various existing solutions to improve torque and power density, a reduction in the dimensions of the end-windings has been explored, aiming to decrease volume, weight, and losses. However, more compact end-windings often lead to complex shapes of the conductors, especially when preformed hairpin windings are considered. The rectangular cross-section of hairpin conductors makes them prone to deviating out of the bending plane during the forming process. This phenomenon, known as torsional–flexural instability, is influenced by the specific aspect ratio of the cross-section dimensions and the bending direction. This study focuses on understanding this instability phenomenon, aiming to identify a potential threshold of the cross-section aspect ratio. The instability makes it difficult to predict the final geometry, potentially compromising the compliance with the geometric tolerances. A finite element model is developed to analyse a single planar bend in a hairpin conductor. Various cross-section dimensions with different aspect ratios are simulated identifying those that experience instability. Moreover, an experimental campaign is conducted to confirm the occurrence of instability by testing the same single planar bending. The experimental data obtained are used to validate the finite element model for the tested dimensions. The aim is to provide designers with a useful tool to select hairpin geometries that are more suitable for the folding process, contributing to successful assembly and improving the overall design process of preformed hairpin conductors.

Tensile behavior of square-twist origami inspired mechanical metamaterials with soft joints and chirality applications

The performance and behavior of origami-inspired mechanical metamaterials are closely related to their structural design and the joints sub equivalent to the crease, in addition to material selection. Current research on square-twist origami focused on the unidirectional formation between its stable states. However, the complete process of squeezing-folding and stretching-unfolding as well as the mechanical behavior under tensile loading have not been thoroughly studied. In this study, square-twist origami mechanical metamaterials (STOMMs) with soft joints are proposed and investigated. The complete process of typical STOMMs from a flat state to folded stable state by extrusion folding and then returning to a flat state by stretch unfolding is explored using the finite element method. The strain energy and deformation characteristics of STOMMs are revealed at eight special stages during the folding, stable state, and unfolding processes. Additionally, the influence of geometric parameters on strain energy, deformation, and tensile behavior is also investigated. Finally, inspired by origami chirality, combinatorial mechanical metamaterials with self-locking/non-self-locking behavior are proposed and validated for their tensile and self-locking behavior. The studies could provide new content for origami-inspired soft joint mechanical metamaterials in terms of self-locking and load-bearing.

‘Main Mechanical Forces‐Chemical Interactions’ Interplay as a Tool to Elucidate Folding Mechanisms

ABSTRACTThe folding of peptides and proteins is rigidly reliant on the ‘chemical information’ carried by the specific amino acid sequence. In this study, three polypeptides (PDBs: 2jof, 1res and 1prv) were investigated as model systems to assess their folded features, thereby enabling further understanding of mechanisms that play a role in regulating the folding process more widely. A novel physico‐chemical approach of analysis is proposed herein, focusing on chemical interactions and their related mechanical forces that we trust are determinant to drive the folding. Through this methodology, we have predicted the conformations adopted by the three polypeptides and compared the outcomes to those experimentally determined, achieving a substantial structural agreement. Molecular dynamic simulations have been carried out to further support our calculations and structural results. Within the three models, we demonstrate that the interaction of each amino acid residue with its neighbour residues is a crucial determinant for the formation of the 3D stable native structures. This article provides initial evidence that the folding occurs by means of mechanical forces developed upon establishing chemical interactions amongst residues, which, in turn, are peculiar to each specific amino acid present in each position of the peptide chain.

Deployment behavior and mechanical property analysis of Kresling origami structure

Kresling origami, a representative variant of origami, exhibits several distinctive features such as multistable states, changeable stiffness, and compression-torsion coupling deformation. The first focus of this study involves examining the mechanical properties of origami structures, specifically, the equivalent modulus and critical buckling stress during the deployment process are evaluated. Subsequently, the collapsibility of the Kresling origami structure is explicated with regard to energy and strain. In the folding process, when the Kresling angle is 0.0135 rad, the maximum torque is 959 Nmm, and the structure will occur snap-through instability, which is the nonlinear critical buckling load of the failure. After that, the torque decreases rapidly, and then gradually increases slowly. The mechanical model is developed and subsequently evaluated in terms of its flexibility and adjustment of stiffness. The present study investigates the buckling modes and vibration properties of Kresling origami structures, as well as their combination structures. For the buckling of Kresling origami structure, the critical buckling load of Euler buckling is 0.023 N and that of local buckling is 0.67 N. In short, the critical buckling load of the column shell is greater than that of the Kresling origami structure, which is due to the small local stiffness of structure.

New insights into folding tape-spring

The tape-spring is widely used in aerospace, mechanical, and civil engineering due to its unique mechanical properties and compact size. This paper aims to establish a new analytical model to more accurately predict the folding behavior of the tape-spring, including the geometric configuration, folding moment versus longitudinal curvature curve, peak moment, and propagation moment. This model comprehensively considers the influence of stretching strain energy and bending strain energy on the total strain energy, and determines the folding behavior of the tape-spring through the minimum energy principle. The proposed analytical model was compared with Wuest’s model, Yee’s model, and Yao’s model. Experiments of tape-springs made from two different materials were utilized for validation. The study shows that the proposed model has the highest prediction accuracy compared with the classical models, with the error of approximately 10%. The reason for the significant difference in the prediction results between the proposed model and Yao’s model was given, and the relationship between stretching strain energy and bending strain energy during the folding process of the tape-spring was analyzed. The proposed model has broader applicability than classical models and provides new insights and solutions for accurate prediction of the folding behavior of the tape-spring.

Hydrogen bonding heterogeneity correlates with protein folding transition state passage time as revealed by data sonification

Protein-protein and protein-water hydrogen bonding interactions play essential roles in the way a protein passes through the transition state during folding or unfolding, but the large number of these interactions in molecular dynamics (MD) simulations makes them difficult to analyze. Here, we introduce a state space representation and associated "rarity" measure to identify and quantify transition state passage (transit) events. Applying this representation to a long MD simulation trajectory that captured multiple folding and unfolding events of the GTT WW domain, a small protein often used as a model for the folding process, we identified three transition categories: Highway (faster), Meander (slower), and Ambiguous (intermediate). We developed data sonification and visualization tools to analyze hydrogen bond dynamics before, during, and after these transition events. By means of these tools, we were able to identify characteristic hydrogen bonding patterns associated with "Highway" versus "Meander" versus "Ambiguous" transitions and to design algorithms that can identify these same folding pathways and critical protein-water interactions directly from the data. Highly cooperative hydrogen bonding can either slow down or speed up transit. Furthermore, an analysis of protein-water hydrogen bond dynamics at the surface of WW domain shows an increase in hydrogen bond lifetime from folded to unfolded conformations with Ambiguous transitions as an outlier. In summary, hydrogen bond dynamics provide a direct window into the heterogeneity of transits, which can vary widely in duration (by a factor of 10) due to a complex energy landscape.

Structure-Foldable and Performance-Tailorable PI Paper-Based Triboelectric Nanogenerators Processed and Controlled by Laser-Induced Graphene.

Laser-induced graphene (LIG) technology has provided a new manufacturing strategy for the rapid and scalable assembling of triboelectric nanogenerators (TENG). However, current LIG-based TENG commonly rely on polymer films, e.g., polyimide (PI) as both friction material and carbon precursor of electrodes, which limit the structural diversity and performance escalation due to its incapability of folding and creasing. Using specialized PI paper composed of randomly distributed PI fibers to substantially enhance its foldability, this work creates a new type of TENG, which are structurally foldable and stackable, and performance tailorable. First, by systematically investigating the laser power-regulated performance of single-unit TENG, the open-circuit voltage can be effectively improved. By further exploiting the folding process, multiple TENG units can be assembled together to form multi-layered structures to continuously expand the open-circuit voltage from 5.3 to 34.4Vcm-2, as the increase of friction units from 1 to 16. Last, by fully utilizing the unique structure and performance, representative energy-harvesting and smart-sensing applications are demonstrated, including a smart shoe to recognize running motions and power LEDs, a smart leaf to power a thermometer by wind, a matrix sensor to recognize writing trajectories, as well as a smart glove to recognize different objects.

Helical reorganization in the context of membrane protein folding: Insights from simulations with bacteriorhodopsin (BR) fragments

Membrane protein folding is distinct from folding of soluble proteins. Conformational acquisition in major membrane protein subclasses can be delineated into insertion and folding processes. An exception to the “two stage” folding, later developed to “three stage” folding, is observed within the last two helices in bacteriorhodopsin (BR), a system that serves as a model membrane protein. We employ a reductionist approach to understand interplay of molecular factors underlying the apparent defiance. Leveraging available solution NMR structures, we construct, sample in silico, and analyze partially (PIn) and fully inserted (FIn) BR membrane states. The membrane lateral C-terminal helix (CH) in PIn is markedly prone to transient structural distortions over microsecond timescales; a disorder prone region (DPR) is thereby identified. While clear transmembrane propensities are not acquired, the distortions induce alterations in local membrane curvature and area per lipid. Importantly, energetic decompositions reveal that overall, the N-terminal helix (NH) is thermodynamically more stable in the PIn. Higher overall stability of the FIn arises from favorable interactions between the NH and the CH. Our results establish lack of spontaneous transition of the PIn to the FIn, and attributes their partitioning to barriers that exceed those accessible with thermal fluctuations. This work paves the way for further detailed studies aimed at determining the thermo-kinetic roles of the initial five helices, or complementary external factors, in complete helical folding and insertion in BR. We comment that complementing such efforts with the growing field of machine learning assisted energy landscape searches may offer unprecedented insights.

Folding Kinetics and Volume Variation of the β-Hairpin Peptide Chignolin upon Ultrafast pH-Jumps.

In-depth characterization of fundamental folding steps of small model peptides is crucial for a better understanding of the folding mechanisms of more complex biomacromolecules. We have previously reported on the folding/unfolding kinetics of a model α-helix. Here, we study folding transitions in chignolin (GYDPETGTWG), a short β-hairpin peptide previously used as a model to study conformational changes in β-sheet proteins. Although previously suggested, until now, the role of the Tyr2-Trp9 interaction in the folding mechanism of chignolin was not clear. In the present work, pH-dependent conformational changes of chignolin were characterized by circular dichroism (CD), nuclear magnetic resonance (NMR), ultrafast pH-jump coupled with time-resolved photoacoustic calorimetry (TR-PAC), and molecular dynamics (MD) simulations. Taken together, our results present a comprehensive view of chignolin's folding kinetics upon local pH changes and the role of the Tyr2-Trp9 interaction in the folding process. CD data show that chignolin's β-hairpin formation displays a pH-dependent skew bell-shaped curve, with a maximum close to pH 6, and a large decrease in β-sheet content at alkaline pH. The β-hairpin structure is mainly stabilized by aromatic interactions between Tyr2 and Trp9 and CH-π interactions between Tyr2 and Pro4. Unfolding of chignolin at high pH demonstrates that protonation of Tyr2 is essential for the stability of the β-hairpin. Refolding studies were triggered by laser-induced pH-jumps and detected by TR-PAC. The refolding of chignolin from high pH, mainly due to the protonation of Tyr2, is characterized by a volume expansion (10.4 mL mol-1), independent of peptide concentration, in the microsecond time range (lifetime of 1.15 μs). At high pH, the presence of the deprotonated hydroxyl (tyrosinate) hinders the formation of the aromatic interaction between Tyr2 and Trp9 resulting in a more disorganized and dynamic tridimensional structure of the peptide. This was also confirmed by comparing MD simulations of chignolin under conditions mimicking neutral and high pH.

Unraveling the Connection: Pain and Endoplasmic Reticulum Stress.

Pain is a complex and multifaceted experience. Recent research has increasingly focused on the role of endoplasmic reticulum (ER) stress in the induction and modulation of pain. The ER is an essential organelle for cells and plays a key role in protein folding and calcium dynamics. Various pathological conditions, such as ischemia, hypoxia, toxic substances, and increased protein production, may disturb protein folding, causing an increase in misfolding proteins in the ER. Such an overload of the folding process leads to ER stress and causes the unfolded protein response (UPR), which increases folding capacity in the ER. Uncompensated ER stress impairs intracellular signaling and cell function, resulting in various diseases, such as diabetes and degenerative neurological diseases. ER stress may be a critical universal mechanism underlying human diseases. Pain sensations involve the central as well as peripheral nervous systems. Several preclinical studies indicate that ER stress in the nervous system is enhanced in various painful states, especially in neuropathic pain conditions. The purpose of this narrative review is to uncover the intricate relationship between ER stress and pain, exploring molecular pathways, implications for various pain conditions, and potential therapeutic strategies.

Utilizing deep ocean water in yeast fermentation for enhanced mineral-rich biomass production and fermentative regulation by proteomics modulation

Deep Ocean Water (DOW) is rich in minerals and serves as a natural source of nutrients. However, due to the inorganic nature of these minerals, cultivating yeast in DOW could aid in the fermentation process, and simultaneously, the yeast can assimilate the minerals from DOW, resulting in a mineral-enriched yeast biomass. Focusing on three DOW sources off the eastern coast of Taiwan (TT-1, HL-1, HL-2), we fermented various yeast strains of Saccharomyces cerevisiae. Therefore, this study investigates the effects of DOW on yeast growth, alcohol dehydrogenase activity, and the biological absorption of mineral ions by the yeast. Additionally, this research employs two-dimensional electrophoresis techniques to examine how the absorbed minerals influence the regulation of yeast proteins, thereby affecting biomass and metabolism. In the result, S. cerevisiae BCRC 21689 demonstrated a remarkable ability to bio-absorb minerals such as magnesium, calcium, potassium, and zinc from DOW, enhancing its growth and fermentation performance. Proteomic analysis revealed significant shifts in the expression of 21 proteins related to glycolytic and energy metabolism, alcohol metabolism, and growth regulation, all influenced by DOW's mineral-rich environment. This indicates that DOW's mineral content is a key factor in upregulating essential enzymes in glycolytic metabolism and alcohol dehydrogenase. An increase in proteins involved in synthesis and folding processes was also observed, leading to a substantial increase in yeast biomass. This study underscores the potential of DOW as a natural enhancer in yeast fermentation processes, enriching the yeast with diverse minerals and modulating proteomic expression to optimize yeast growth and fermentation.

Sortase A-Based Post-translational Modifications on Encapsulin Nanocompartments.

Protein-based encapsulin nanocompartments, known for their well-defined structures and versatile functionalities, present promising opportunities in the fields of biotechnology and nanomedicine. In this investigation, we effectively developed a sortase A-mediated protein ligation system in Escherichia coli to site-specifically attach target proteins to encapsulin, both internally and on its surfaces without any further in vitro steps. We explored the potential applications of fusing sortase enzyme and a protease for post-translational ligation of encapsulin to a green fluorescent protein and anti-CD3 scFv. Our results demonstrated that this system could attach other proteins to the nanoparticles' exterior surfaces without adversely affecting their folding and assembly processes. Additionally, this system enabled the attachment of proteins inside encapsulins which varied shapes and sizes of the nanoparticles due to cargo overload. This research developed an alternative enzymatic ligation method for engineering encapsulin nanoparticles to facilitate the conjugation process.

The Folding Pathway of ABC Transporter CFTR: Effective and Robust

De novo protein folding into a native three-dimensional structure is indispensable for biological function, is instructed by its amino acid sequence, and occurs along a vectorial trajectory. The human proteome contains thousands of membrane-spanning proteins, whose biosynthesis begins on endoplasmic reticulum-associated ribosomes. Nearly half of all membrane proteins traverse the membrane more than once, including therapeutically important protein families such as solute carriers, G-protein-coupled receptors, and ABC transporters. These mediate a variety of functions like signal transduction and solute transport and are often of vital importance for cell function and tissue homeostasis. Missense mutations in multispan membrane proteins can lead to misfolding and cause disease; an example is the ABC transporter Cystic Fibrosis Transmembrane Conductance Regulator (CFTR). Even though our understanding of multispan membrane-protein folding still is rather rudimental, the cumulative knowledge of 20 years of basic research on CFTR folding has led to development of drugs that modulate the misfolded protein. This has provided the prospect of a life without CF to the vast majority of patients. In this review we describe our understanding of the folding pathway of CFTR in cells, which is modular and tolerates many defects, making it effective and robust. We address how modulator drugs affect folding and function of CFTR, and distinguish protein stability from its folding process. Since the domain architecture of (mammalian) ABC transporters are highly conserved, we anticipate that the insights we discuss here for folding of CFTR may lay the groundwork for understanding the general rules of ABC-transporter folding.

Numerical modelling of different airbag folding patterns and their influence on occupant responses in frontal vehicle impact

This paper presented a procedure for airbag folding for the application in occupant safety studies and analysed the influence of different airbag folding patterns on the occupant severity in frontal impact. Airbags were folded in two patterns: zig-zag and top-roll, using two folding techniques: Initial Metric Method and Explicit Folding. The explicit folding was found to be more expensive in terms of preparation time. However, this approach provided more control over the whole folding process. The Initial Metric Method was more robust, however, it’s hard to apply for complex folding patterns. Deployments of airbags were validated against experimental data of pendulum tests. Those airbag models were applied to 2006 Ford F250 pickup truck. This vehicle was used for simulations of frontal vehicle impact where the 50th male Hybrid III crash test dummy was an occupant. Results of this simulation were compared with an actual test with a similar vehicle, under the same impact conditions. Results showed the head and chest accelerations were lower for top-roll folded airbag cases, however neck normal forces were higher compared to zig-zag folded airbag. The internal pressure in the early stage of deployment was 33% higher for the top-roll folded airbags.

Terapi Bermain Melipat dan Menempel Kertas Origami Pada Anak Usia Dini

Introduction: The benefits of origami for young children are numerous. First, origami can help develop children's fine motor skills. The process of folding and gluing paper requires good hand-eye coordination, as well as fine motor skills. Second, origami can also help develop children's creativity and imagination. Children can create various shapes and objects just by folding paper. Method: This play therapy activity was carried out on Friday 17 November 2023 at the Al-Hidayah Ciomas Islamic Education Foundation, Panjalu sub-district. Participants in this activity were 24 preschool children and RA Al-Hidayah. The method used in this activity is divided into three stages, namely the preparation, implementation and final evaluation stages. In the preparation stage, we prepare tools, make time contracts, convey goals and topics, practice together, and prepare prizes for participants who have good work. Folded paper and hvs are used as media used by children to express their creativity. The instructions given to children are not limited, in the sense that children can express any ideas on hvs paper with origami tools as the main material. The goal is for students to have high thinking and creativity in creating creations using origami paper. Finally, in the evaluation stage, the staff reviews and assesses the children's work on how they draw using origami paper as the main basic material. Result: Playing activities for children were carried out on Friday 17 November 2023 at the Al-Hidayah Ciomas Islamic Education Foundation, Panjalu sub-district, this activity was attended by 29 children as participants. It is known that the implementation of play activities for children runs smoothly. Which was opened by the event section and continued by other implementing sections until the stage of implementing therapy for participants. And continued with evaluation, closing and documentation.Conclusion: Play therapy through origami, the Japanese art of paper folding, is an effective method for early childhood development. It helps in developing fine motor skills, creativity, imagination, and focus.

Origami metamaterials for ultra-wideband and large-depth reflection modulation

The dynamic control of electromagnetic waves is a persistent pursuit in modern industrial development. The state-of-the-art dynamic devices suffer from limitations such as narrow bandwidth, limited modulation range, and expensive features. To address these issues, we fuse origami techniques with metamaterial design to achieve ultra-wideband and large-depth reflection modulation. Through a folding process, our proposed metamaterial achieves over 10-dB modulation depth over 4.96 – 38.8 GHz, with a fractional bandwidth of 155% and tolerance to incident angles and polarizations. Its ultra-wideband and large-depth reflection modulation performance is verified through experiments and analyzed through multipole decomposition theory. To enhance its practical applicability, transparent conductive films are introduced to the metamaterial, achieving high optical transparency (>87%) from visible to near-infrared light while maintaining cost-effectiveness. Benefiting from lightweight, foldability, and low-cost properties, our design shows promise for extensive satellite communication and optical window mobile communication management.

A systemic approach to identifying sequence frameworks that decrease mAb production in a transient Chinese hamster ovary cell expression system.

Monoclonal antibodies (mAbs) are often engineered at the sequence level for improved clinical performance yet are rarely evaluated prior to candidate selection for their "developability" characteristics, namely expression, which can necessitate additional resource investments to improve the manufacturing processes for problematic mAbs. A strong relationship between primary sequence and expression has emerged, with slight differences in amino acid sequence resulting in titers differing by up to an order of magnitude. Previous work on these "difficult-to-express" (DTE) mAbs has shown that these phenotypes are driven by post-translational bottlenecks in antibody folding, assembly, and secretion processes. However, it has been difficult to translate these findings across cell lines and products. This work presents a systematic approach to study the impact of sequence variation on mAb expression at a larger scale and under more industrially relevant conditions. The analysis found 91 mutations that decreased transient expression of an IgG1κ in Chinese hamster ovary (CHO) cells and revealed that mutations at inaccessible residues, especially those leading to decreases in residue hydrophobicity, are not favorable for high expression. This workflow can be used to better understand sequence determinants of mAb expression to improve candidate selection procedures and reduce process development timelines.

Transcriptomic WGCNA analyses reveal endoplasmic reticulum response of Patinopecten yessoensis under acute heat stress

Patinopecten yessoensis is one of the most important aquaculture shellfish worldwide. As a cold-water species, the increasing seawater temperatures due to global warming have been a great challenge for P. yessoensis. Understanding the response mechanism of P. yessoensis to high temperatures is essential to the genetic breeding of heat-tolerant varieties. In this study, histological, cytological and transcriptomic analyses were performed on the gill tissues of P. yessoensis under different acute heat stresses (temperature: 22 °C, 25 °C, 28 °C; time: 6 h, 36 h). A total of 10,601 differentially expressed genes (DEGs) were detected, among which, 141 DEGs were shared by all heat-stressed groups. Weighted gene co-expression network analysis (WGCNA) identified three modules (M1, M2 and M3) that were significantly correlated with heat stress. Functional enrichment analysis revealed that heat stress probably caused serious endoplasmic reticulum (ER) stress in gill cells, with many associated GO terms and KEGG pathways markedly enriched. Notably, the protein processing in ER pathway probably played a key role in the response to heat stress; this pathway was upregulated and was the most enriched in all heat-stressed groups, as well as in the M1 and M2 modules. DEGs from this pathway participated in different processes of the ER stress response, alleviating ER stress by upregulating the protein correct folding, ER-associated degradation (ERAD) and unfolded protein response (UPR) processes. Excessive ER stress induced apoptosis via the UPR, and excessive apoptosis resulted in tissue injury as observed by histological and TUNEL assays. Further, numerous Hsps were upregulated in all heat-stressed groups, and some members were identified as hub genes, including BiP, NEF, Hsp70 and Hsp90. These genes may perform crucial functions during heat stress as molecular chaperones. This study elucidates the key molecular components of the ER stress response to heat stress in scallops. The findings provide new insights into the adaptation mechanisms of molluscs to environmental stress.

DNA Damage Accelerates G-Quadruplex Folding in a Duplex-G-Quadruplex-Duplex Context.

Molecular details for the impact of DNA damage on folding of potential G-quadruplex sequences (PQSs) to noncanonical DNA structures involved in gene regulation are poorly understood. Here, the effects of DNA base damage and strand breaks on PQS folding kinetics were studied in the context of the VEGF promoter sequence embedded between two DNA duplex anchors, termed a duplex-G-quadruplex-duplex (DGD) motif. This DGD scaffold imposes constraints on the PQS folding process that more closely mimic those found in genomic DNA. Folding kinetics were monitored by circular dichroism (CD) to find folding half-lives ranging from 2 s to 12 min depending on the DNA damage type and sequence position. The presence of Mg2+ ions and G-quadruplex (G4)-binding protein APE1 facilitated the folding reactions. A strand break placing all four G runs required for G4 formation on one side of the break accelerated the folding rate by >150-fold compared to the undamaged sequence. Combined 1D 1H NMR and CD analyses confirmed that isothermal folding of the VEGF-DGD constructs yielded spectral signatures that suggest the formation of G4 motifs and demonstrated a folding dependency on the nature and location of DNA damage. Importantly, the PQS folding half-lives measured are relevant to replication, transcription, and DNA repair time frames.

Mechanism of DNA origami folding elucidated by mesoscopic simulations

Many experimental and computational efforts have sought to understand DNA origami folding, but the time and length scales of this process pose significant challenges. Here, we present a mesoscopic model that uses a switchable force field to capture the behavior of single- and double-stranded DNA motifs and transitions between them, allowing us to simulate the folding of DNA origami up to several kilobases in size. Brownian dynamics simulations of small structures reveal a hierarchical folding process involving zipping into a partially folded precursor followed by crystallization into the final structure. We elucidate the effects of various design choices on folding order and kinetics. Larger structures are found to exhibit heterogeneous staple incorporation kinetics and frequent trapping in metastable states, as opposed to more accessible structures which exhibit first-order kinetics and virtually defect-free folding. This model opens an avenue to better understand and design DNA nanostructures for improved yield and folding performance.