Fold Structures Research Articles

Clinical and genetic analysis of essential hypertension with CYB gene m.15024G>A mutation.

To explore the role of mitochondrial CYB 15024G>A mutation in the development of essential hypertension. Mitochondrial genome sequences of hypertensive patients were obtained from previous studies. Clinical and genetic data of a hypertensive patient with mitochondrial CYB 15024G>A mutation and its pedigree were analyzed. Lymphocytes derived from patient and family members were transformed into immortalized lymphoblastoid cell lines, and the levels of adenosine triphosphate (ATP), mitochondrial membrane potential and intracellular reactive oxygen species (ROS) were detected. The penetrance of this essential hypertension family was 42.9%, and the age of onset was 46-68 years old. Mitochondrial genome sequencing results showed that all maternal members carried a highly conserved mitochondrial CYB 15024G>A mutation. This mutation could affect the free energy of mitochondrial CYB for secondary and tertiary structure and protein folding, thereby changing its structural stability and the structure of the electron transfer function area around the mutation site. Compared with the control, the cell line carrying the mitochondrial CYB 15024G>A mutation showed significantly decreased levels of mitochondrial CYB, ATP and mitochondrial membrane potential, and increased levels of ROS (P<0.01). Mitochondrial CYB 15024G>A mutation may affect the structure of respiratory chain subunits and mitochondrial function, leading to cell dysfunction, which suggests that the mutation may play a synergistic role in essential hypertension.

The handling of tunnel collapse and settlement (case study: Jragung Dam diversion tunnel, Indonesia)

A diversion tunnel is a crucial part of dam construction because of its function to derivate the river during earth dam filling works. The Jragung Dam diversion tunnel is located in the Kalibeng Formation, which is dominated by claystone and sandstone lithology, and passed by an anticline fold structure. The geological conditions provide challenges in tunnel construction. The collapse of rock material in the tunnel overburden and settlement on the slopes hill requires a comprehensive approach and handling by prioritizing factors of safety, cost efficiency, and speed of time. Field observation including rock mass quality analysis, reviewing the design, also monitoring of discharge and deformation was conducted to find the source of collapse. The groundwater flow and deformed soft rock were accused of being the tunnel collapse determinant. The rock mass improvements used pore-filling of sodium silicate mixture and fore-polling. Then dewatering was carried out using drainage pipes and deep wells. Because of the steep rock’s dip in the tunnel, the release wells and observation wells location need to design effectively. Geoelectric investigations correlated with bore log analysis aimed to find effective locations for groundwater control. The settlement was refilled with plastic concrete because of its lightweight and durability.

Mono-valent salt corrections for RNA secondary structures in the ViennaRNA package

BackgroundRNA features a highly negatively charged phosphate backbone that attracts a cloud of counter-ions that reduce the electrostatic repulsion in a concentration dependent manner. Ion concentrations thus have a large influence on folding and stability of RNA structures. Despite their well-documented effects, salt effects are not handled consistently by currently available secondary structure prediction algorithms. Combining Debye-Hückel potentials for line charges and Manning’s counter-ion condensation theory, Einert et al. (Biophys J 100: 2745-2753, 2011) modeled the energetic contributions of monovalent cations on loops and helices.ResultsThe model of Einert et al. is adapted to match the structure of the dynamic programming recursion of RNA secondary structure prediction algorithms. An empirical term describing the salt dependence of the duplex initiation energy is added to improve co-folding predictions for two or more RNA strands. The slightly modified model is implemented in the ViennaRNA package in such way that only the energy parameters but not the algorithmic structure is affected. A comparison with data from the literature show that predicted free energies and melting temperatures are in reasonable agreement with experiments.ConclusionThe new feature in the ViennaRNA package makes it possible to study effects of salt concentrations on RNA folding in a systematic manner. Strictly speaking, the model pertains only to mono-valent cations, and thus covers the most important parameter, i.e., the NaCl concentration. It remains a question for future research to what extent unspecific effects of bi- and tri-valent cations can be approximated in a similar manner.AvailabilityCorrections for the concentration of monovalent cations are available in the ViennaRNA package starting from version 2.6.0.

An onshore-offshore interpretation of structures in the Devonian rocks of the Pentland Firth, Scotland using high resolution bathymetry and drone-enabled field observations.

Strong tidal currents in the Pentland Firth separating NE Scotland and Orkney have stripped recent seafloor sediments revealing geological structures in parts of the Devonian Orcadian Basin that are otherwise limited to narrow coastal outcrops. Here we interpret and analyse a 220 km2 high-resolution bathymetric dataset and combine these findings with onshore aerial imagery, field observations, and photogrammetric Virtual Outcrop Models created from coastal outcrops. This approach allows a reappraisal of the structure, stratigraphy and tectonic evolution of the Orcadian Basin, and gives new insights into fold and fault structures developed at sub-seismic to reservoir scales. A major basin-scale Devonian structure – the Brough-Brims-Risa Fault – can be traced from Caithness to Hoy and has partitioned reactivation-related deformation in the Orcadian Basin during later deformation episodes. Carboniferous inversion-related folds formed during E-W shortening are mapped and correlated between Caithness and Orkney. These structures are cross-cut by widespread highly connected fracture networks formed by steeply-dipping ENE-WSW and subordinate WNW-ESE trending structures developed during Permian NW-SE transtensional rifting. The offshore dataset demonstrates the continuity and topology of structures related to superimposed rifting and basin inversion events across a much greater scale range than was hitherto possible. [end].

Unveiling the functional epitopes of cobra venom cytotoxin by immunoinformatics and epitope-omic analyses

Approximate 70% of cobra venom is composed of cytotoxin (CTX), which is responsible for the dermonecrotic symptoms of cobra envenomation. However, CTX is generally low in immunogenicity, and the antivenom is ineffective in attenuating its in vivo toxicity. Furthermore, little is known about its epitope properties for empirical antivenom therapy. This study aimed to determine the epitope sequences of CTX using the immunoinformatic analyses and epitope-omics profiling. A conserved CTX was used in this study to determine its T-cell and B-cell epitope sequences using immunoinformatic tools and molecular docking simulation with different Human Leukocyte Antigens (HLAs). The potential T-cell and B-cell epitopes were 'KLVPLFY,' 'CPAGKNLCY,' 'MFMVSTPTK,' and 'DVCPKNSLL.' Molecular docking simulations disclosed that the HLA-B62 supertype exhibited the greatest binding affinity towards cobra venom cytotoxin. The namely L7, G18, K19, N20, M25, K33, V43, C44, K46, N47, and S48 of CTX exhibited prominent intermolecular interactions with HLA-B62. The multi-enzymatic-limited-digestion/liquid chromatography-mass spectrometry (MELD/LC–MS) also revealed three potential epitope sequences as 'LVPLFYK,' 'MFMVS,' and ‘TVPVKR’. From different epitope mapping approaches, we concluded four potential epitope sites of CTX as ‘KLVPLFYK’, ‘AGKNL’, ‘MFMVSTPKVPV’ and ‘DVCPKNSLL’. Site-directed mutagenesis of these epitopes confirmed their locations at the functional loops of CTX. These epitope sequences are crucial to CTX’s structural folding and cytotoxicity. The results concluded the epitopes that resided within the functional loops constituted potential targets to fabricate synthetic epitopes for CTX-targeted antivenom production.

Efficient Hi-C inversion facilitates chromatin folding mechanism discovery and structure prediction

Genome-wide chromosome conformation capture (Hi-C) experiments have revealed many structural features of chromatin across multiple length scales. Further understanding genome organization requires relating these discoveries to the mechanisms that establish chromatin structures and reconstructing these structures in three dimensions, but both objectives are difficult to achieve with existing algorithms that are often computationally expensive. To alleviate this challenge, we present an algorithm that efficiently converts Hi-C data into contact energies, which measure the interaction strength between genomic loci brought into proximity. Contact energies are local quantities unaffected by the topological constraints that correlate Hi-C contact probabilities. Thus, extracting contact energies from Hi-C contact probabilities distills the biologically unique information contained in the data. We show that contact energies reveal the location of chromatin loop anchors, support a phase separation mechanism for genome compartmentalization, and parameterize polymer simulations that predict three-dimensional chromatin structures. Therefore, we anticipate that contact energy extraction will unleash the full potential of Hi-C data and that our inversion algorithm will facilitate the widespread adoption of contact energy analysis.

Genomic and Temporal Analysis of Deletions Correlated to qRT-PCR Dropout in N Gene in Alpha, Delta and Omicron Variants.

Since the first SARS-CoV-2 outbreak, mutations such as single nucleotide polymorphisms (SNPs) and insertion/deletions (INDELs) have changed and characterized the viral genome sequence, structure and protein folding leading to the onset of new variants. The presence of those alterations challenges not only the clinical field but also the diagnostic demand due to failures in gene detection or incompleteness of polymerase chain reaction (PCR) results. In particular, the analysis of understudied genes such as N and the investigation through whole-genome next generation sequencing (WG-NGS) of regions more prone to mutate can help in the identification of new or reacquired mutations, with the aim of designing robust and long-lasting primers. In 48 samples of SARS-CoV-2 (including Alpha, Delta and Omicron variants), a lack of N gene amplification was observed in the genomes analyzed through WG-NGS. Three gene regions were detected hosting the highest number of SNPs and INDELs. In several cases, the latter can interfere deeply with both the sensitivity of diagnostic methodologies and the final protein folding. The monitoring over time of the viral evolution and the reacquisition among different variants of the same mutations or different alterations within the same genomic positions can be relevant to avoid unnecessary consumption of resources.

Three-Dimensional Geological–Geophysical Modeling and Prospecting Indications of the Ashele Ore Concentration Area in Xinjiang Based on Irregular Sections

The Ashele ore concentration area is an important area for polymetallic ore concentration in Xinjiang, China. Scholars have made progress in understanding the ore-controlling structures, ore-bearing horizons, and metallogenic age of this area. However, there are still uncertainties about the 3D distributions of plutons, fault structures, and ore-bearing strata, which restrict the development of deep and peripheral ore prospecting and the discovery of new ore bodies in the area. This study proposes a geological–geophysical modeling method based on irregular sections and uses this method to establish a 3D geological–geophysical model based on physical property data, boreholes, surface geological maps, and geophysical data. The model shows that the study area has many hidden rock masses with various depths and shapes and fracture structures with complex shapes. The fault structure in the area is complex, and the ore bodies are controlled by the faults. The ore-bearing geological units (Ashele Formation) exhibit an obvious east–west-trending W-shaped fold structure. The deep part of the northern Ashele Formation extends northward slightly, and the southern Ashele Formation has thick strata, with depths generally greater than 2 km. Based on the information on deep structures provided by the model, three metallogenic prospective areas are predicted, which points out the direction for further prospecting work in the ore concentration area and shows that the adopted modeling method and process have good applicability for constructing 3D models of ore concentration areas with sparse data, large area, and complex geological structures. The proposed modeling method provides technical support for ore prospecting, particularly in the overburden area or ore concentration area with sparse data.

State-of-the-art and novel approaches to mild solubilization of inclusion bodies.

Throughout the twenty-first century, the view on inclusion bodies (IBs) has shifted from undesired by-products towards a targeted production strategy for recombinant proteins. Inclusion bodies can easily be separated from the crude extract after cell lysis and contain the product in high purity. However, additional solubilization and refolding steps are required in the processing of IBs to recover the native protein. These unit operations remain a highly empirical field of research in which processes are developed on a case-by-case basis using elaborate screening strategies. It has been shown that a reduction in denaturant concentration during protein solubilization can increase the subsequent refolding yield due to the preservation of correctly folded protein structures. Therefore, many novel solubilization techniques have been developed in the pursuit of mild solubilization conditions that avoid total protein denaturation. In this respect, ionic liquids have been investigated as promising agents, being able to solubilize amyloid-like aggregates and stabilize correctly folded protein structures at the same time. This review briefly summarizes the state-of-the-art of mild solubilization of IBs and highlights some challenges that prevent these novel techniques from being yet adopted in industry. We suggest mechanistic models based on the thermodynamics of protein unfolding with the aid of molecular dynamics simulations as a possible approach to solve these challenges in the future.

Micro-CT driven characterization and validation of constituents’ properties used in origami-inspired foldable composites fabricated via different manufacturing processes

High packaging efficient lightweight jointless deployable folding structures are nowadays fabricated from combination of rigid and thin flexible elastic hinges using fiber reinforced polymers composites (FRPCs) for various applications such as space, robotics, flexible electronics, etc. It is of high importance to optimize these structures in the design phase using computer aided analysis tools for which the structural-level elastic constants are challenging to find experimentally. In this work, a generic X-ray computed tomography (micro-CT) driven framework based on voxel modeling approach was developed to virtually characterize the smallest building block of a typical large-scale foldable structure that consists of a rigid region and elastic hinge needed for folding operation. The voxel-based description of a real representative volume element (RVE) was generated using the images obtained from the micro-CT. The homogenized mechanical properties for each of the material systems was then obtained by multi-parameter component segmentation and subsequent directional averaging of the corresponding voxels. Three-dimensional void analyses were also shown using the micro-CT based voxel modeling. The accuracy of the non-destructive framework was demonstrated for four materials systems, based on different combinations of the matrix and fabric architectures in comparison with the mechanical tests. The homogenized mechanical properties obtained using the present framework based on the real RVE showed good agreement with the experimental results providing a foundation for a structural-level computer aided analysis.

A New design of deployable pedestrian bridge deck based on origami theory

As one of the basic civil engineering structures, bridges greatly impact people’s daily life. However, many bridge accidents are caused by boat crashes because of the driver’s carelessness or other climatic and hydrological factors. Besides, some pedestrian bridges are not frequently used and thus make a waste of water space. This design will overcome the challenge and tackle the problem successfully based on the origami and kirigami theory by using a connector composed of a slider and a revolute joint. The bridge can be folded up and down via a particular folding method. The basic structure units have two degrees of freedom (DOF). The whole mechanism can be folded and unfolded by sliding and rotating the connector. According to the structural folding position analysis results shown in the diagrams, the new deployable bridge deck structure design can validly tackle the problems of clearance limitation under the bridge via the special-designed cross-section of the bridge deck. I hope the design model will be widely applied in the river-crossing bridge and bring convenience to all pedestrians and boat drivers.

Protein–Protein Interfaces as Druggable Targets: A Common Motif of the Pyridoxal-5′-Phosphate-Dependent Enzymes to Receive the Coenzyme from Its Producers

Pyridoxal-5′-phosphate (PLP), a phosphorylated form of vitamin B6, acts as a coenzyme for numerous reactions, including those changed in cancer and/or associated with the disease prognosis. Since highly reactive PLP can modify cellular proteins, it is hypothesized to be directly transferred from its donors to acceptors. Our goal is to validate the hypothesis by finding common motif(s) in the multitude of PLP-dependent enzymes for binding the limited number of PLP donors, namely pyridoxal kinase (PdxK), pyridox(am)in-5′-phosphate oxidase (PNPO), and PLP-binding protein (PLPBP). Experimentally confirmed interactions between the PLP donors and acceptors reveal that PdxK and PNPO interact with the most abundant PLP acceptors belonging to structural folds I and II, while PLPBP – with those belonging to folds III and V. Aligning sequences and 3D structures of the identified interactors of PdxK and PNPO, we have identified a common motif in the PLP-dependent enzymes of folds I and II. The motif extends from the enzyme surface to the neighborhood of the PLP binding site, represented by an exposed alfa-helix, a partially buried beta-strand, and residual loops. Pathogenicity of mutations in the human PLP-dependent enzymes within or in the vicinity of the motif, but outside of the active sites, supports functional significance of the motif that may provide an interface for the direct transfer of PLP from the sites of its synthesis to those of coenzyme binding. The enzyme-specific amino acid residues of the common motif may be useful to develop selective inhibitors blocking PLP delivery to the PLP-dependent enzymes critical for proliferation of malignant cells.

Design of a 2RRU-RRS Parallel Kinematic Mechanism for an Inner-Cavity Machining Hybrid Robot

Abstract This article presents a hybrid robot for machining the inner cavity of large-scale workpiece, and it is composed of three parts: a mobile device with 1T degree-of-freedom (DoF), a serial module with 1R1T DoFs, and a 2RRU-RRS parallel kinematic mechanism (PKM) with 2R1T DoFs. The 2RRU-RRS PKM has some advantages with a folding structure, only one S joint, two certain rotational axes, and all the fixed actuators. In this article, the conceptual design, theoretical kinematic and dynamic modeling, performance evaluation, and optimization of the parallel system are investigated. A 3D printing model is built to demonstrate the application potential. This article plays an exemplary role in the design of inner-cavity machining hybrid robots.

Normal faulting and landsliding in morpho-structural domes related to buried salt stocks, Zagros Mountains, Iran. Insights into salt breakout

Normal faulting in domal structures above diapirs has important implications in hydrocarbon geoscience, but studies on surface faulting on exposed growing domes are scarce. Moreover, there is limited work addressing the erosional processes involved in the unroofing of active diapirs preceding salt emergence. This investigation analyses two morpho-structural domes (Gavbast Dome: GD; Kooh Bedoo Dome: KBD) associated with growing anticlines related to the rejuvenation of precursor buried salt stocks in the Zagros Mountains. The structural relief of the domes (1330 m in GD and 400 m in KBD) and the available chronological data on fold development indicate long-term uplift rates of around 0.1–0.4 mm/yr. GD displays three domains with different fault patterns: (1) fold-normal grabens in one half; (2) polygonal faulting with multiple enclosed graben depressions in the crest; and (3) fold-parallel faulting on the other half. KBD shows a major fold-normal graben restricted to the crest and one flank of the dome. The lack of the typical radial pattern related to multidirectional hoop extension can be attributed the suppression of extension in the fold-normal direction by regional shortening. Other factors such as complexities in the folding structure, outer-arc extension in the anticline crests, and plan geometry seem to play a local control on fault patterns. The studied domes display an extraordinarily high density of large slope movements, indicating that they play an instrumental role in the unroofing process. The slope failures, with sliding planes mainly developed in argillaceous units overlain by thick limestone packages, include incipient planar rock slides more than 5 km2 in area expressed as buckle folds in the lower part of the slopes. The main preparatory factors involved in the development of these slope collapses include rock mass weakening by faulting and slope over-steepening, both related to doming. Our detailed mapping and the examination of a number of salt diapirs in the Fars Arc suggest that episodic landsliding and more gradual fluvial dissection, often controlled by grabens, are the main erosional processes involved in the unroofing of active diapirs leading to salt emergence.

Impact of Rehydration Following Systemic Dehydration on Vocal Fold Gene Expression.

Biological data on the beneficial effects of vocal fold rehydration are lacking. This study aimed to examine the effects of acute systemic dehydration on vocal fold gene expression and determine whether rehydration would reverse these changes. Male New Zealand White rabbits (N = 24, n = 8/group) provided the animal model. Systemic dehydration was induced by 5 days of water volume restriction. Rehydration was provided by ad-lib water for 3 days following dehydration. Euhydrated rabbits were used as the control group. Vocal fold tissue was dissected. Seventeen genes were selected based on physiological function and role in supporting vocal fold structure, oxidative stress, hemodynamics, and extracellular matrix turnover. Relative gene expression was assessed by RT-qPCR. Rehydration following systemic dehydration can modulate gene expression, with expression patterns suggesting that rehydration reverses dehydration-induced changes in over half of the tested genes. CLIC5 (chloride intracellular channel 5) and EFEMP1 (EGF containing fibulin extracellular matrix protein 1) genes were significantly upregulated in the dehydration group compared with the euhydrated control. A1BG (alpha-1B-glycoprotein) and IL1RAP (interleukin 1 receptor accessory protein) were downregulated by rehydration compared with the dehydration group. This study provides molecular evidence for a transcriptional response to rehydration following acute systemic dehydration in the vocal folds. These data are the first to study gene expression following realistic dehydration and rehydration paradigms and provide biological data to support clinical recommendations to increase water intake after acute dehydration. NA Laryngoscope, 133:3499-3505, 2023.

Computational Modeling of DNA 3D Structures: From Dynamics and Mechanics to Folding.

DNA carries the genetic information required for the synthesis of RNA and proteins and plays an important role in many processes of biological development. Understanding the three-dimensional (3D) structures and dynamics of DNA is crucial for understanding their biological functions and guiding the development of novel materials. In this review, we discuss the recent advancements in computer methods for studying DNA 3D structures. This includes molecular dynamics simulations to analyze DNA dynamics, flexibility, and ion binding. We also explore various coarse-grained models used for DNA structure prediction or folding, along with fragment assembly methods for constructing DNA 3D structures. Furthermore, we also discuss the advantages and disadvantages of these methods and highlight their differences.

GraphGPSM: a global scoring model for protein structure using graph neural networks.

The scoring models used for protein structure modeling and ranking are mainly divided into unified field and protein-specific scoring functions. Although protein structure prediction has made tremendous progress since CASP14, the modeling accuracy still cannot meet the requirements to a certain extent. Especially, accurate modeling of multi-domain and orphan proteins remains a challenge. Therefore, an accurate and efficient protein scoring model should be developed urgently to guide the protein structure folding or ranking through deep learning. In this work, we propose a protein structure global scoring model based on equivariant graph neural network (EGNN), named GraphGPSM, to guide protein structure modeling and ranking. We construct an EGNN architecture, and a message passing mechanism is designed to update and transmit information between nodes and edges of the graph. Finally, the global score of the protein model is output through a multilayer perceptron. Residue-level ultrafast shape recognition is used to describe the relationship between residues and the overall structure topology, and distance and direction encoded by Gaussian radial basis functions are designed to represent the overall topology of the protein backbone. These two features are combined with Rosetta energy terms, backbone dihedral angles and inter-residue distance and orientations to represent the protein model and embedded into the nodes and edges of the graph neural network. The experimental results on the CASP13, CASP14 and CAMEO test sets show that the scores of our developed GraphGPSM have a strong correlation with the TM-score of the models, which are significantly better than those of the unified field score function REF2015 and the state-of-the-art local lDDT-based scoring models ModFOLD8, ProQ3D and DeepAccNet, etc. The modeling experimental results on 484 test proteins demonstrate that GraphGPSM can greatly improve the modeling accuracy. GraphGPSM is further used to model 35 orphan proteins and 57 multi-domain proteins. The results show that the average TM-score of the models predicted by GraphGPSM is 13.2 and 7.1% higher than that of the models predicted by AlphaFold2. GraphGPSM also participates in CASP15 and achieves competitive performance in global accuracy estimation.

Structure and ion-dependent folding of k-junctions.

k-Junctions are elaborated forms of kink turns with an additional helix on the nonbulged strand, thus forming a three-way helical junction. Two were originally identified in the structures of Arabidopsis and Escherichia coli thiamine pyrophosphate (TPP) riboswitches, and another called DUF-3268 was tentatively identified from sequence information. In this work we show that the Arabidopsis and E. coli riboswitch k-junctions fold in response to the addition of magnesium or sodium ions, and that atomic mutations that should disrupt key hydrogen bonding interactions greatly impair folding. Using X-ray crystallography, we have determined the structure of the DUF-3268 RNA and thus confirmed that it is a k-junction. It also folds upon the addition of metal ions, though requiring a 40-fold lower concentration of either divalent or monovalent ions. The key difference between the DUF-3268 and riboswitch k-junctions is the lack of nucleotides inserted between G1b and A2b in the former. We show that this insertion is primarily responsible for the difference in folding properties. Finally, we show that the DUF-3268 can functionally substitute for the k-junction in the E. coli TPP riboswitch such that the chimera can bind the TPP ligand, although less avidly.

Study on stress evolution law and rock burst mechanism in upright fold structure area of deep mine

With the increase in coal mining depth, many rock bursts triggered by fold structures have been observed, but the disaster mechanism is not clear. This research aims at investigating the influence of upright fold structure(UFS) on rock burst mechanisms. The mechanical model and numerical model of the UFS are constructed, and the mining stress evolution law and rock burst mechanism of working face in a UFS area are studied by utilizing engineering practice. The research shows that the occurrence of rock burst disasters in the fold structure area has obvious regional characteristics. The horizontal and vertical stress of the UFS in the horizontal distribution is similar to the periodic variation characteristics of the sine (cosine) curve. The stress state of the UFS area is divided into five areas. The horizontal and vertical stress concentration factors increase first and then decrease with the working face gradually approaching and away from the upright fold syncline axis. A roadway impact risk index I 0 is proposed to determine whether the roadway is damaged, and the mechanism of inducing ground pressure during mining in UFS areas of deep mines is expounded. The control plan effectively reduces the stress levels of surrounding rock in working faces.

Bioinformatics characterization of BcsA-like orphan proteins suggest they form a novel family of pseudomonad cyclic-β-glucan synthases.

Bacteria produce a variety of polysaccharides with functional roles in cell surface coating, surface and host interactions, and biofilms. We have identified an 'Orphan' bacterial cellulose synthase catalytic subunit (BcsA)-like protein found in four model pseudomonads, P. aeruginosa PA01, P. fluorescens SBW25, P. putida KT2440 and P. syringae pv. tomato DC3000. Pairwise alignments indicated that the Orphan and BcsA proteins shared less than 41% sequence identity suggesting they may not have the same structural folds or function. We identified 112 Orphans among soil and plant-associated pseudomonads as well as in phytopathogenic and human opportunistic pathogenic strains. The wide distribution of these highly conserved proteins suggest they form a novel family of synthases producing a different polysaccharide. In silico analysis, including sequence comparisons, secondary structure and topology predictions, and protein structural modelling, revealed a two-domain transmembrane ovoid-like structure for the Orphan protein with a periplasmic glycosyl hydrolase family GH17 domain linked via a transmembrane region to a cytoplasmic glycosyltransferase family GT2 domain. We suggest the GT2 domain synthesises β-(1,3)-glucan that is transferred to the GH17 domain where it is cleaved and cyclised to produce cyclic-β-(1,3)-glucan (CβG). Our structural models are consistent with enzymatic characterisation and recent molecular simulations of the PaPA01 and PpKT2440 GH17 domains. It also provides a functional explanation linking PaPAK and PaPA14 Orphan (also known as NdvB) transposon mutants with CβG production and biofilm-associated antibiotic resistance. Importantly, cyclic glucans are also involved in osmoregulation, plant infection and induced systemic suppression, and our findings suggest this novel family of CβG synthases may provide similar range of adaptive responses for pseudomonads.