Structural analysis of a trimeric assembly of the mitochondrial dynamin-like GTPase Mgm1.

The fusion of inner mitochondrial membranes requires dynamin-like GTPases, Mgm1 in yeast and OPA1 in mammals, but how they mediate membrane fusion is poorly understood. Here, we determined the crystal structure of Saccharomyces cerevisiae short Mgm1 (s-Mgm1) in complex with GDP. It revealed an N-terminal GTPase (G) domain followed by two helix bundles (HB1 and HB2) and a unique C-terminal lipid-interacting stalk (LIS). Dimers can form through antiparallel HB interactions. Head-to-tail trimers are built by intermolecular interactions between the G domain and HB2-LIS. Biochemical and in vivo analyses support the idea that the assembly interfaces observed here are native and critical for Mgm1 function. We also found that s-Mgm1 interacts with negatively charged lipids via both the G domain and LIS. Based on these observations, we propose that membrane targeting via the G domain and LIS facilitates the in cis assembly of Mgm1, potentially generating a highly curved membrane tip to allow inner membrane fusion

Multi-Label Takagi-Sugeno-Kang Fuzzy System

Multi-label classification can effectively identify the relevant labels of an instance from a given set of labels. However,the modeling of the relationship between the features and the labels is critical to the classification performance. To this end, we propose a new multi-label classification method, called Multi-Label Takagi-Sugeno-Kang Fuzzy System (ML-TSK FS), to improve the classification performance. The structure of ML-TSK FS is designed using fuzzy rules to model the relationship between features and labels. The fuzzy system is trained by integrating fuzzy inference based multi-label correlation learning with multi-label regression loss. The proposed ML-TSK FS is evaluated experimentally on 12 benchmark multi-label datasets. 1 The results show that the performance of ML-TSK FS is competitive with existing methods in terms of various evaluation metrics, indicating that it is able to model the feature-label relationship effectively using fuzzy inference rules and enhances the classification performance.Comment: This work has been accepted by IEEE Transactions on Fuzzy System

Four years of multi-modal odometry and mapping on the rail vehicles

Precise, seamless, and efficient train localization as well as long-term railway environment monitoring is the essential property towards reliability, availability, maintainability, and safety (RAMS) engineering for railroad systems. Simultaneous localization and mapping (SLAM) is right at the core of solving the two problems concurrently. In this end, we propose a high-performance and versatile multi-modal framework in this paper, targeted for the odometry and mapping task for various rail vehicles. Our system is built atop an inertial-centric state estimator that tightly couples light detection and ranging (LiDAR), visual, optionally satellite navigation and map-based localization information with the convenience and extendibility of loosely coupled methods. The inertial sensors IMU and wheel encoder are treated as the primary sensor, which achieves the observations from subsystems to constrain the accelerometer and gyroscope biases. Compared to point-only LiDAR-inertial methods, our approach leverages more geometry information by introducing both track plane and electric power pillars into state estimation. The Visual-inertial subsystem also utilizes the environmental structure information by employing both lines and points. Besides, the method is capable of handling sensor failures by automatic reconfiguration bypassing failure modules. Our proposed method has been extensively tested in the long-during railway environments over four years, including general-speed, high-speed and metro, both passenger and freight traffic are investigated. Further, we aim to share, in an open way, the experience, problems, and successes of our group with the robotics community so that those that work in such environments can avoid these errors. In this view, we open source some of the datasets to benefit the research community

Determination of Fluoroquinolone Residues in Aquatic Products by Dispersive Solid Phase Extraction with Metal-Organic Framework-Based Molecularly Imprinted Polymer and Liquid Chromatography-Tandem Mass Spectrometry

A novel adsorption material for dispersive solid phase extraction (DSPE) was prepared using amino-functionalized zirconium-based metal-organic framework (UiO-66-NH2) as the core and molecularly imprinted polymer as the shell. Subsequently, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) were used to characterize the material. Key experiential parameters such as solvent, adsorbent concentration, adsorption time, desorption solvent and time were systematically investigated. Furthermore, a highly sensitive method for the determination of fluoroquinolone residues in aquatic products was developed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with DSPE. The extract was blown to near dryness under nitrogen and re-dissolved with 10% methanol-ammonia solution (pH 8.0). Subsequently, 30 mg of the adsorbent was added for adsorption for 8 minutes. Next, the fluoroquinolones sorbed were ultrasonically desorbed with 10% acetic acid-methanol solution for 4 minutes, blown to near dryness under nitrogen and re-dissolved before analysis by LC-MS/MS. Good linearity was observed all 10 fluoroquinolones in the concentration range of 0.1–200 μg/L, with recovery rates of 84.5%–105.9%. Compared with the national standard method, the established method had higher accuracy, lower limit of detection (LOD) and limit of quantification (LOQ), simpler and more efficient operation. The new material had good selectivity and reusability, effectively reducing the cost of detection. This study helps expand the application scope of metal-organic framework-based molecularly imprinted polymers in food safety detection

Crystal Structure of the C-Terminal Cytoplasmic Domain of Non-Structural Protein 4 from Mouse Hepatitis Virus A59

BACKGROUND:The replication of coronaviruses takes place on cytoplasmic double membrane vesicles (DMVs) originating in the endoplasmic reticulum (ER). Three trans-membrane non-structural proteins, nsp3, nsp4 and nsp6, are understood to be membrane anchors of the coronavirus replication complex. Nsp4 is localized to the ER membrane when expressed alone but is recruited into the replication complex in infected cells. It is revealed to contain four trans-membrane regions and its N- and C-termini are exposed to the cytosol. METHODOLOGY/PRINCIPAL FINDINGS:We have determined the crystal structures of the C-terminal hydrophilic domain of nsp4 (nsp4C) from MHV strain A59 and a C425S site-directed mutant. The highly conserved 89 amino acid region from T408 to Q496 is shown to possess a new fold. The wild-type (WT) structure features two monomers linked by a Cys425-Cys425 disulfide bond in one asymmetric unit. The monomers are arranged with their N- and C-termini in opposite orientations to form an "open" conformation. Mutation of Cys425 to Ser did not affect the monomer structure, although the mutant dimer adopts strikingly different conformations by crystal packing, with the cross-linked C-termini and parallel N-termini of two monomers forming a "closed" conformation. The WT nsp4C exists as a dimer in solution and can dissociate easily into monomers in a reducing environment. CONCLUSIONS/SIGNIFICANCE:As nsp4C is exposed in the reducing cytosol, the monomer of nsp4C should be physiological. This structure may serve as a basis for further functional studies of nsp4

Structural Basis of Enzymatic Activity for the Ferulic Acid Decarboxylase (FADase) from Enterobacter sp. Px6-4

Microbial ferulic acid decarboxylase (FADase) catalyzes the transformation of ferulic acid to 4-hydroxy-3-methoxystyrene (4-vinylguaiacol) via non-oxidative decarboxylation. Here we report the crystal structures of the Enterobacter sp. Px6-4 FADase and the enzyme in complex with substrate analogues. Our analyses revealed that FADase possessed a half-opened bottom β-barrel with the catalytic pocket located between the middle of the core β-barrel and the helical bottom. Its structure shared a high degree of similarity with members of the phenolic acid decarboxylase (PAD) superfamily. Structural analysis revealed that FADase catalyzed reactions by an “open-closed” mechanism involving a pocket of 8×8×15 Å dimension on the surface of the enzyme. The active pocket could directly contact the solvent and allow the substrate to enter when induced by substrate analogues. Site-directed mutagenesis showed that the E134A mutation decreased the enzyme activity by more than 60%, and Y21A and Y27A mutations abolished the enzyme activity completely. The combined structural and mutagenesis results suggest that during decarboxylation of ferulic acid by FADase, Trp25 and Tyr27 are required for the entering and proper orientation of the substrate while Glu134 and Asn23 participate in proton transfer